195 lines
11 KiB
Plaintext
195 lines
11 KiB
Plaintext
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
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polymerase"
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CDS complement(140..157)
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/codon_start=1
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/label=6xHis
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/note="6xHis affinity tag"
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/translation="HHHHHH"
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CDS complement(207..239)
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/codon_start=1
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/label=T7 tag (gene 10 leader)
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/note="leader peptide from bacteriophage T7 gene 10"
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/translation="MASMTGGQQMG"
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CDS complement(243..260)
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/codon_start=1
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/label=thrombin site
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/note="thrombin recognition and cleavage site"
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/translation="LVPRGS"
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CDS complement(270..287)
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/codon_start=1
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/label=6xHis
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/note="6xHis affinity tag"
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/translation="HHHHHH"
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RBS complement(306..328)
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/label=RBS
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/note="efficient ribosome binding site from bacteriophage
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T7 gene 10 (Olins and Rangwala, 1989)"
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protein_bind complement(343..367)
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/label=lac operator
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/note="The lac repressor binds to the lac operator to
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inhibit transcription in E. coli. This inhibition can be
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relieved by adding lactose or
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isopropyl-beta-D-thiogalactopyranoside (IPTG)."
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promoter complement(368..386)
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/label=T7 promoter
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/note="promoter for bacteriophage T7 RNA polymerase"
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promoter 695..772
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/label=lacI promoter
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CDS 773..1852
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/codon_start=1
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/label=lacI
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/note="lac repressor"
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/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
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NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
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EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
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EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
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MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
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YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
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ALADSLMQLARQVSRLESGQ"
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protein_bind 1868..1889
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/label=CAP binding site
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/note="CAP binding activates transcription in the presence
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of cAMP."
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CDS 2664..2852
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/codon_start=1
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/label=rop
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/note="Rop protein, which maintains plasmids at low copy
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number"
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/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
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DELYRSCLARFGDDGENL"
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misc_feature 2957..3099
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/label=bom
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/note="basis of mobility region from pBR322"
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rep_origin complement(3285..3873)
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/direction=LEFT
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/label=ori
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/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
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replication"
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CDS 3995..4807
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/codon_start=1
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/label=KanR
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/note="aminoglycoside phosphotransferase"
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/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
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KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
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TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
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SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
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ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
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rep_origin complement(4903..5358)
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/direction=LEFT
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/label=f1 ori
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/note="f1 bacteriophage origin of replication; arrow
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indicates direction of (+) strand synthesis"
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ORIGIN
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1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
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61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
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121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
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181 cgacggagct cgaattcgga tccgcgaccc atttgctgtc caccagtcat gctagccata
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241 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg
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301 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
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361 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
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421 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
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481 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
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541 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
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601 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
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661 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
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721 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
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781 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
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841 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
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901 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
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961 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
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1021 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
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1081 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
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1141 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
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1201 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
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1261 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
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1321 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
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1381 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
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1441 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
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1501 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
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1561 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
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1621 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
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1681 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
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1741 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
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1801 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
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1861 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
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1921 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
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1981 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
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2041 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
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2101 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
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2161 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
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2221 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
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2281 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
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2341 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
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2401 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
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2461 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
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2521 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
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2581 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
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2641 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
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2701 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
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2761 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
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2821 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
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2881 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
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2941 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
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3001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa
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3061 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
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3121 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
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3181 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
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3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
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3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
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3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
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3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
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3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
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3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
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3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
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3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
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3721 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
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3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
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3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
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3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact
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3961 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
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4021 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
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4081 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
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4141 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
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4201 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
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4261 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
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4321 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
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4381 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
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4441 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
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4501 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
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4561 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
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4621 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
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4681 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
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4741 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
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4801 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac
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4861 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
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4921 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
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4981 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
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5041 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
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5101 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
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5161 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
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5221 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
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5281 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
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5341 cgccgctaca gggcgcgtcc cattcgcca
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//
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