add gfp2
This commit is contained in:
mpabi
parent
68367f04c3
commit
4f3350fd7a
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#! /usr/bin/env python3
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# vim:fenc=utf-8
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#
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# Copyright © 2024 user <user@penguin>
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#
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# Distributed under terms of the MIT license.
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import os
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from Bio import Entrez, SeqIO
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from Bio.Seq import Seq
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from Bio.SeqFeature import SeqFeature, FeatureLocation
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from Bio.Restriction import RestrictionBatch, EcoRI, BamHI, HindIII, Bpu1102I
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from Bio.Restriction import AllEnzymes, Analysis
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from Bio.Graphics import GenomeDiagram
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from reportlab.lib import colors
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#%%
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# Ustawienia Entrez
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Entrez.email = "your_email@example.com" # Wpisz swój adres email
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# Funkcja do pobierania sekwencji z NCBI i zapisywania jej lokalnie
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def fetch_and_save_sequence(db, id, file_path):
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handle = Entrez.efetch(db=db, id=id, rettype="gb", retmode="text")
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record = SeqIO.read(handle, "genbank")
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handle.close()
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SeqIO.write(record, file_path, "genbank")
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return record
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# Funkcja do wczytywania sekwencji z lokalnego pliku
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def load_local_sequence(file_path):
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with open(file_path, 'r') as file:
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record = SeqIO.read(file, "genbank")
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return record
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#%%
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# ID dla sekwencji w NCBI
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gfp_id = "U87974"
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plasmid_id = "pET-28a(+)"
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# Åcieżki do lokalnych plików
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gfp_file_path = gfp_id+'.gb'
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plasmid_file_path = plasmid_id+'.gb'
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#%%
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# Sprawdzanie i pobieranie sekwencji GFP
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if not os.path.exists(gfp_file_path):
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print(f"Plik {gfp_file_path} nie istnieje. Pobieranie z serwera NCBI...")
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gfp_record = fetch_and_save_sequence("nuccore", gfp_id, gfp_file_path)
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print(f"Pobrano i zapisano sekwencjÄ GFP do pliku {gfp_file_path}.")
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else:
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print(f"Plik {gfp_file_path} istnieje. Wczytywanie danych z lokalnego pliku...")
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gfp_record = load_local_sequence(gfp_file_path)
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print(f"Wczytano dane z pliku {gfp_file_path}.")
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gfp_seq = str(gfp_record.seq)
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print(f"GFP Sequence: {gfp_seq[:60]}...")
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#%%
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# Sprawdzanie i pobieranie sekwencji plazmidu
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if not os.path.exists(plasmid_file_path):
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print(f"Plik {plasmid_file_path} nie istnieje. Pobieranie z serwera NCBI...")
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record = fetch_and_save_sequence("nuccore", plasmid_id, plasmid_file_path)
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print(f"Pobrano i zapisano sekwencjÄ plazmidu do pliku {plasmid_file_path}.")
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else:
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print(f"Plik {plasmid_file_path} istnieje. Wczytywanie danych z lokalnego pliku...")
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record = load_local_sequence(plasmid_file_path)
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print(f"Wczytano dane z pliku {plasmid_file_path}.")
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#%%
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# WyÅwietl etykiety (annotacje)
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for feature in record.features:
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print(f"Type: {feature.type}")
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print(f"Location: {feature.location}")
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if 'gene' in feature.qualifiers:
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print(f"Gene: {feature.qualifiers['gene']}")
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if 'product' in feature.qualifiers:
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print(f"Product: {feature.qualifiers['product']}")
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print()
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#%%
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#%%
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# Przejdź przez wszystkie funkcje w rekordzie i wypisz ich szczegóÅowe informacje
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for feature in record.features:
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if feature.type in ["promoter", "RBS","CDS", "protein_bind", "misc_feature", "rep_origin", "terminator"]:
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print(f"Type: {feature.type}")
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print(f"Location: {feature.location}")
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print(f"Strand: {'+' if feature.strand == 1 else '-'}")
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# SzczegóÅowe opisy
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if feature.type == "CDS":
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gene_name = feature.qualifiers.get('gene', ['Unknown gene'])[0]
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product = feature.qualifiers.get('product', ['Unknown product'])[0]
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print(f"Gene Name: {gene_name}")
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print(f"Product: {product}")
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elif feature.type == "protein_bind":
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binding_site = feature.qualifiers.get('bound_moiety', ['Unknown binding site'])[0]
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print(f"Binding Site: {binding_site}")
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elif feature.type == "misc_feature":
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note = feature.qualifiers.get('note', ['No additional information'])[0]
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print(f"Note: {note}")
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elif feature.type == "rep_origin":
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print("This is a replication origin.")
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# Pobierz sekwencjÄ funkcji
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feature_seq = record.seq[feature.location.start:feature.location.end]
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print(f"Sequence ( ): {feature_seq}\n")
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# UwzglÄdnij orientacjÄ nici (strand)
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if feature.strand == -1:
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feature_seq = feature_seq.reverse_complement()
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print(f"Sequence (^): {feature_seq}\n")
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# Znajdź i wyÅwietl sekwencjÄ terminatora T7
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for feature in record.features:
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if feature.type == "terminator" and "T7" in feature.qualifiers.get('note', [''])[0]:
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print("Terminator T7 znaleziony:")
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print(f"Type: {feature.type}")
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print(f"Location: {feature.location}")
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print(f"Sequence: {record.seq[feature.location.start:feature.location.end]}")
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print()
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#%%
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# Znajdź i wyÅwietl sekwencjÄ terminatora T7
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for feature in record.features:
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if feature.type == "terminator" and "T7" in feature.qualifiers.get('note', [''])[0]:
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print("Terminator T7 znaleziony:")
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print(f"Type: {feature.type}")
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print(f"Location: {feature.location}")
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# Pobierz sekwencjÄ terminatora
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terminator_seq = record.seq[feature.location.start:feature.location.end]
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# Sprawdź orientacjÄ i odwróÄ/uzupeÅnij jeÅli potrzebne
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if feature.strand == -1:
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terminator_seq = terminator_seq.reverse_complement()
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print(f"(+) Sequence: {terminator_seq}")
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print()
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#%%
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#%%
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t=record[25:73].seq
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#%%
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a = str (t)
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a
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#%%
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class Model:
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complement = {'A': 'T', 'T': 'A', 'C': 'G', 'G': 'C'}
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def __init__ (self, seq=None):
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self.seq = seq
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def complement_dna(self):
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complementary_sequence = ''.join(self.complement[base] for base in self.seq)
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return complementary_sequence
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#%%
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r = Model(a)
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#%%
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r.seq
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#%%
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r.seq[::-1]
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#%%
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r.complement_dna()
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#%%
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r.seq
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#%%
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complement = {'A': 'T', 'T': 'A', 'C': 'G', 'G': 'C'}
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complementary_sequence = ''.join(complement[base] for base in r.seq)
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complementary_sequence
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#%%
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#%%
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# Sekwencja DNA
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dna_seq = record.seq
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# Znajdź miejsca ciÄcia dla NcoI i Bpu1102I
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ncoI_sites = NcoI.search(dna_seq)
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bpu1102I_sites = Bpu1102I.search(dna_seq)
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# Przyjmij pierwsze miejsca ciÄcia
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ncoI_site = ncoI_sites[0]
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bpu1102I_site = bpu1102I_sites[0]
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# WyodrÄbnij fragmenty 20 zasad od miejsc ciÄcia
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ncoI_fragment = dna_seq[ncoI_site-16:ncoI_site+4] # 16 zasad przed i 4 po ciÄciu
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bpu1102I_fragment = dna_seq[bpu1102I_site:bpu1102I_site+20] # 20 zasad po ciÄciu
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# Lepkie koÅce
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ncoI_sticky_end = ncoI_fragment[-4:]
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ncoI_sticky_end_comp = ncoI_sticky_end.complement()
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bpu1102I_sticky_end = bpu1102I_fragment[:4]
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bpu1102I_sticky_end_comp = bpu1102I_sticky_end.complement()
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# Funkcja do wyÅwietlania dwuniciowego DNA z lepkimi koÅcami w formacie schodkowym z indeksami dla dÅuższej nici
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def print_sticky_ends(seq1, sticky_end1, seq2, sticky_end2, start1, start2):
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# Convert sequences to strings
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seq1_str = str(seq1)
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sticky_end1_str = str(sticky_end1)
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comp_seq1_str = str(seq1.complement())
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sticky_end1_comp_str = str(sticky_end1.complement())
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seq2_str = str(seq2)
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sticky_end2_str = str(sticky_end2)
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comp_seq2_str = str(seq2.complement())
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sticky_end2_comp_str = str(sticky_end2.complement())
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# Add indexes to the ends
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seq1_with_index = f"{seq1_str[:-1]} -{start1+len(seq1_str)-1}"
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comp_seq1_with_index = f"{comp_seq1_str[:-1]}"
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seq2_with_index = f"{start2}- {seq2_str[1:]}"
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comp_seq2_with_index = f"{comp_seq2_str[1:]}"
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# Lustrzane odbicie sekwencji
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comp_seq2_with_index = comp_seq2_with_index[::-1]
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sticky_end2_comp_str = sticky_end2_comp_str[::-1]
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# Format the output as requested
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print(f"5'-{seq1_with_index} {sticky_end1_str}-3' (+)")
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print(f"3'-{comp_seq1_with_index}{sticky_end1_comp_str}-5'")
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print()
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print(f"5'-{sticky_end2_str} {seq2_with_index}-3' (+)")
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print(f" {sticky_end2_comp_str}{comp_seq2_with_index}-5'")
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# WyÅwietl lepkie koÅce fragmentów
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print("Fragmenty po ciÄciu NcoI i Bpu1102I:")
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print_sticky_ends(ncoI_fragment[:-4], ncoI_sticky_end, bpu1102I_fragment[4:], bpu1102I_sticky_end, ncoI_site-16, bpu1102I_site)
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#%%
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# Projektowanie starterów
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forward_primer = gfp_seq[:20] # Pierwsze 20 nukleotydów sekwencji GFP
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reverse_primer = str(Seq(gfp_seq_with_his6[-20:]).reverse_complement()) # Ostatnie 20 nukleotydów + His6
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print(f"Forward Primer: {forward_primer}")
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print(f"Reverse Primer: {reverse_primer}")
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#%%
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# Miejsca ciÄcia restryktazy
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enzymes = RestrictionBatch([EcoRI, BamHI, HindIII])
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#enzymes = RestrictionBatch([BamHI, ])
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restriction_sites = enzymes.search(record.seq)
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#%%
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# Przeprowadzenie analizy restrykcyjnej
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analysis = Analysis(AllEnzymes, record.seq)
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# Wynik analizy
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results = analysis.full()
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# WyÅwietlanie wyników
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#for enzyme in results:
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# if len(results[enzyme]) > 0:
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# print(f"{enzyme}: {results[enzyme]}")
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#%%
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# Utwórz diagram plazmidu
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diagram = GenomeDiagram.Diagram("PUC19 Plasmid Map")
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track = diagram.new_track(1, name="Annotated Features", greytrack=True)
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feature_set = track.new_set()
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# Dodanie sekwencji kodujÄ
cej GFP
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feature_set.add_feature(
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SeqFeature(FeatureLocation(0, len(gfp_seq)), strand=+1),
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name="GFP Coding Sequence",
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label=True,
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color=colors.lightblue
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)
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# Dodanie His6 tagu
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feature_set.add_feature(
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SeqFeature(FeatureLocation(len(gfp_seq), len(gfp_seq_with_his6)), strand=+1),
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name="His6 Tag",
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label=True,
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color=colors.lightgreen
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)
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# Dodanie forward primer
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feature_set.add_feature(
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SeqFeature(FeatureLocation(0, len(forward_primer)), strand=+1),
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name="Forward Primer",
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label=True,
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color=colors.orange
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)
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# Dodanie reverse primer
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feature_set.add_feature(
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SeqFeature(FeatureLocation(len(gfp_seq_with_his6) - len(reverse_primer), len(gfp_seq_with_his6)), strand=-1),
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name="Reverse Primer",
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label=True,
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color=colors.red
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)
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# Dodanie miejsc ciÄcia restryktazy
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for enzyme, sites in restriction_sites.items():
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for site in sites:
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feature_set.add_feature(
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SeqFeature(FeatureLocation(site, site + 1), strand=0),
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name=enzyme,
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label=True,
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color=colors.purple
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)
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# Rysowanie diagramu
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diagram.draw(format="circular", circular=True, pagesize='A4', start=0, end=len(record), circle_core=0.5)
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diagram.write("pdf/plasmid_map.pdf", "PDF")
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print("Zapisano diagram plazmidu do pliku 'plasmid_map.pdf'")
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@ -0,0 +1,194 @@
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LOCUS 40924_17796 5369 bp DNA circular SYN 14-OCT-2021
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DEFINITION synthetic circular DNA.
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ACCESSION .
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VERSION .
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KEYWORDS .
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SOURCE synthetic DNA construct
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ORGANISM synthetic DNA construct
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REFERENCE 1 (bases 1 to 5369)
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AUTHORS caoheibi
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TITLE Direct Submission
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REFERENCE 2 (bases 1 to 5369)
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AUTHORS .
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TITLE Direct Submission
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COMMENT SGRef: number: 1; type: "Journal Article"
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FEATURES Location/Qualifiers
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source 1..5369
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/mol_type="other DNA"
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/organism="synthetic DNA construct"
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terminator complement(26..73)
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/label=T7 terminator
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|
/note="transcription terminator for bacteriophage T7 RNA
|
||||||
|
polymerase"
|
||||||
|
CDS complement(140..157)
|
||||||
|
/codon_start=1
|
||||||
|
/label=6xHis
|
||||||
|
/note="6xHis affinity tag"
|
||||||
|
/translation="HHHHHH"
|
||||||
|
CDS complement(207..239)
|
||||||
|
/codon_start=1
|
||||||
|
/label=T7 tag (gene 10 leader)
|
||||||
|
/note="leader peptide from bacteriophage T7 gene 10"
|
||||||
|
/translation="MASMTGGQQMG"
|
||||||
|
CDS complement(243..260)
|
||||||
|
/codon_start=1
|
||||||
|
/label=thrombin site
|
||||||
|
/note="thrombin recognition and cleavage site"
|
||||||
|
/translation="LVPRGS"
|
||||||
|
CDS complement(270..287)
|
||||||
|
/codon_start=1
|
||||||
|
/label=6xHis
|
||||||
|
/note="6xHis affinity tag"
|
||||||
|
/translation="HHHHHH"
|
||||||
|
RBS complement(306..328)
|
||||||
|
/label=RBS
|
||||||
|
/note="efficient ribosome binding site from bacteriophage
|
||||||
|
T7 gene 10 (Olins and Rangwala, 1989)"
|
||||||
|
protein_bind complement(343..367)
|
||||||
|
/label=lac operator
|
||||||
|
/note="The lac repressor binds to the lac operator to
|
||||||
|
inhibit transcription in E. coli. This inhibition can be
|
||||||
|
relieved by adding lactose or
|
||||||
|
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
|
||||||
|
promoter complement(368..386)
|
||||||
|
/label=T7 promoter
|
||||||
|
/note="promoter for bacteriophage T7 RNA polymerase"
|
||||||
|
promoter 695..772
|
||||||
|
/label=lacI promoter
|
||||||
|
CDS 773..1852
|
||||||
|
/codon_start=1
|
||||||
|
/label=lacI
|
||||||
|
/note="lac repressor"
|
||||||
|
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
|
||||||
|
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
|
||||||
|
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
|
||||||
|
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
|
||||||
|
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
|
||||||
|
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
|
||||||
|
ALADSLMQLARQVSRLESGQ"
|
||||||
|
protein_bind 1868..1889
|
||||||
|
/label=CAP binding site
|
||||||
|
/note="CAP binding activates transcription in the presence
|
||||||
|
of cAMP."
|
||||||
|
CDS 2664..2852
|
||||||
|
/codon_start=1
|
||||||
|
/label=rop
|
||||||
|
/note="Rop protein, which maintains plasmids at low copy
|
||||||
|
number"
|
||||||
|
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
|
||||||
|
DELYRSCLARFGDDGENL"
|
||||||
|
misc_feature 2957..3099
|
||||||
|
/label=bom
|
||||||
|
/note="basis of mobility region from pBR322"
|
||||||
|
rep_origin complement(3285..3873)
|
||||||
|
/direction=LEFT
|
||||||
|
/label=ori
|
||||||
|
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
|
||||||
|
replication"
|
||||||
|
CDS 3995..4807
|
||||||
|
/codon_start=1
|
||||||
|
/label=KanR
|
||||||
|
/note="aminoglycoside phosphotransferase"
|
||||||
|
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
|
||||||
|
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
|
||||||
|
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
|
||||||
|
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
|
||||||
|
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
|
||||||
|
rep_origin complement(4903..5358)
|
||||||
|
/direction=LEFT
|
||||||
|
/label=f1 ori
|
||||||
|
/note="f1 bacteriophage origin of replication; arrow
|
||||||
|
indicates direction of (+) strand synthesis"
|
||||||
|
ORIGIN
|
||||||
|
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
|
||||||
|
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
|
||||||
|
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
|
||||||
|
181 cgacggagct cgaattcgga tccgcgaccc atttgctgtc caccagtcat gctagccata
|
||||||
|
241 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg
|
||||||
|
301 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
|
||||||
|
361 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
|
||||||
|
421 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
|
||||||
|
481 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
|
||||||
|
541 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
|
||||||
|
601 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
|
||||||
|
661 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
|
||||||
|
721 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
|
||||||
|
781 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
|
||||||
|
841 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
|
||||||
|
901 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
|
||||||
|
961 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
|
||||||
|
1021 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
|
||||||
|
1081 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
|
||||||
|
1141 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
|
||||||
|
1201 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
|
||||||
|
1261 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
|
||||||
|
1321 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
|
||||||
|
1381 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
|
||||||
|
1441 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
|
||||||
|
1501 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
|
||||||
|
1561 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
|
||||||
|
1621 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
|
||||||
|
1681 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
|
||||||
|
1741 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
|
||||||
|
1801 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
|
||||||
|
1861 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
|
||||||
|
1921 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
|
||||||
|
1981 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
|
||||||
|
2041 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
|
||||||
|
2101 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
|
||||||
|
2161 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
|
||||||
|
2221 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
|
||||||
|
2281 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
|
||||||
|
2341 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
|
||||||
|
2401 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
|
||||||
|
2461 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
|
||||||
|
2521 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
|
||||||
|
2581 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
|
||||||
|
2641 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
|
||||||
|
2701 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
|
||||||
|
2761 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
|
||||||
|
2821 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
|
||||||
|
2881 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
|
||||||
|
2941 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
|
||||||
|
3001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa
|
||||||
|
3061 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
|
||||||
|
3121 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
|
||||||
|
3181 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
|
||||||
|
3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
|
||||||
|
3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
|
||||||
|
3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
|
||||||
|
3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
|
||||||
|
3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
|
||||||
|
3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
|
||||||
|
3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
|
||||||
|
3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
|
||||||
|
3721 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
|
||||||
|
3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
|
||||||
|
3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
|
||||||
|
3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact
|
||||||
|
3961 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
|
||||||
|
4021 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
|
||||||
|
4081 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
|
||||||
|
4141 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
|
||||||
|
4201 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
|
||||||
|
4261 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
|
||||||
|
4321 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
|
||||||
|
4381 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
|
||||||
|
4441 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
|
||||||
|
4501 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
|
||||||
|
4561 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
|
||||||
|
4621 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
|
||||||
|
4681 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
|
||||||
|
4741 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
|
||||||
|
4801 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac
|
||||||
|
4861 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
|
||||||
|
4921 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
|
||||||
|
4981 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
|
||||||
|
5041 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
|
||||||
|
5101 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
|
||||||
|
5161 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
|
||||||
|
5221 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
|
||||||
|
5281 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
|
||||||
|
5341 cgccgctaca gggcgcgtcc cattcgcca
|
||||||
|
//
|
||||||
|
|
||||||
Loading…
Reference in New Issue