LOCUS 40924_17796 5369 bp DNA circular SYN 14-OCT-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5369) AUTHORS caoheibi TITLE Direct Submission REFERENCE 2 (bases 1 to 5369) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5369 /mol_type="other DNA" /organism="synthetic DNA construct" terminator complement(26..73) /label=T7 terminator /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 //