LOCUS . 5711 bp DNA circular UNK 01-JAN-1980 DEFINITION . ACCESSION VERSION KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers promoter complement(464..482) /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff" /label="T7 promoter" protein_bind 439..463 /bound_moiety="lac repressor encoded by lacI" /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-β-D-thiogalactopyranoside (IPTG)." /note="color: #31849b" /label="lac operator" CDS complement(360..389) /codon_start=1 /product="10xHis affinity tag" /transl_table=1 /translation="HHHHHHHHHH" /label="10xHis" /note="color: #cc99b2" terminator 213..260 /note="transcription terminator for bacteriophage T7 RNA polymerase" /note="color: #ffffff" /label="T7 terminator" promoter 791..868 /gene="lacI" /note="" /note="color: #ffffff" /label="lacI promoter" promoter complement(5504..5608) /gene="bla" /label="AmpR promoter" /note="color: #ffffff" CDS 3263..3454 /codon_start=1 /gene="rop" /product="Rop protein" /transl_table=1 /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL*" /label="rop" /note="color: #993366" rep_origin complement(3884..4472) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" /label="ori" CDS complement(4643..5503) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="color: #ccffcc" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" CDS 869..1951 /codon_start=1 /gene="lacI" /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-β-D-thiogalactopyranoside (IPTG)." /note="color: #993366" /product="lac repressor" /transl_table=1 /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ*" /label="lacI" CDS complement(336..347) /codon_start=1 /product="Factor Xa recognition and cleavage site" /transl_table=1 /translation="IEGR" /label="Factor Xa site" /note="color: #cc99b2" CDS complement(393..395) /codon_start=1 /product="start codon" /transl_table=1 /translation="M" /label="ATG" /note="color: #993366" RBS 403..407 /note="ribosome binding site" /note="color: #a6acb3" /label="RBS" misc_feature 319..335 /note="multiple cloning site" /note="color: #99ccff" /label="MCS" ORIGIN 1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa 61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg 121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt 181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag 241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt 301 cgggctttgt tagcagccgg atcctcgagc atatgacgac cttcgatatg gccgctgctg 361 tgatgatgat gatgatgatg atgatgatgg cccatggtat atctccttct taaagttaaa 421 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat 481 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc 541 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg 601 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg 661 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc 721 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt 781 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc 841 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga 901 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc 961 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg 1021 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct 1081 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg 1141 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt 1201 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca 1261 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc 1321 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt 1381 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc 1441 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca 1501 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca 1561 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct 1621 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt 1681 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca 1741 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca 1801 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc 1861 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg 1921 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca 1981 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct 2041 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc 2101 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct 2161 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc 2221 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg 2281 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg 2341 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca 2401 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc 2461 ttaccagcct aacttcgatc actggaccgc tgatcgtcac ggcgatttat gccgcctcgg 2521 cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt gtctgcctcc 2581 ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca 2641 cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt 2701 gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg 2761 cacgcggcgc atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct 2821 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc 2881 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac 2941 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg 3001 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac 3061 atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat 3121 ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt 3181 aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa 3241 tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg 3301 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga 3361 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct 3421 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac 3481 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt 3541 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg 3601 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg cggtgtgaaa 3661 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca 3721 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 3781 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 3841 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 3901 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 3961 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4021 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 4081 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 4141 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 4201 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 4261 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 4321 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 4381 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 4441 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 4501 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 4561 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 4621 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 4681 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 4741 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 4801 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 4861 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 4921 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct 4981 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 5041 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 5101 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 5161 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 5221 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac 5281 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 5341 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 5401 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 5461 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 5521 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 5581 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct 5641 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc 5701 gtcttcaaga a //