LOCUS . 3621 bp DNA circular UNK 01-JAN-1980 DEFINITION . ACCESSION VERSION KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers CDS complement(2399..3481) /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(520..1311) /codon_start=1 /gene="aadA" /note="confers resistance to spectinomycin and streptomycin" /note="color: #ccffcc" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /transl_table=1 /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK*" /label="SmR" rep_origin complement(1451..2189) /direction=LEFT /note="Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." /note="color: #ffff00" /label="CloDF13 ori" promoter complement(1312..1403) /gene="bla" /label="AmpR promoter" /note="color: #ffffff" promoter complement(3482..3559) /gene="lacI" /note="" /note="color: #ffffff" /label="lacI promoter" terminator 302..349 /note="transcription terminator for bacteriophage T7 RNA polymerase" /note="color: #ffffff" /label="T7 terminator" CDS 206..250 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /transl_table=1 /translation="KETAAAKFERQHMDS" /label="S-Tag" /note="color: #cc99b2" protein_bind 3..27 /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" promoter 2..3605 /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff" /label="T7 promoter" CDS 77..94 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" /note="color: #cc99b2" CDS 113..127 /codon_start=1 /product="enterokinase recognition and cleavage site" /transl_table=1 /translation="DDDDK" /label="enterokinase site" /note="color: #cc99b2" RBS 58..63 /note="ribosome binding site" /note="color: #a6acb3" /label="RBS" CDS 71..73 /codon_start=1 /product="start codon" /transl_table=1 /translation="M" /label="ATG" /note="color: #993366" misc_feature 69..278 /note="multiple cloning site" /note="color: #99ccff" /label="MCS" ORIGIN 1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag 61 gagatatacc atggcacatc accaccacca tcacgtgggt accggttcga atgatgacga 121 cgacaagagt ccggatccca attgggagct cgtgtacacg gcgcgcctgc aggtcgacaa 181 gcttgcggcc gcactcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca 241 catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg ctgagcaata 301 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca 361 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca 421 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc 481 ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt 541 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg 601 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc 661 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac 721 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc 781 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg 841 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc 901 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag 961 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg 1021 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc 1081 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag 1141 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac 1201 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc 1261 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct 1321 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 1381 atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag 1441 actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg 1501 gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc 1561 cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg 1621 ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt 1681 tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct 1741 gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc 1801 tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc 1861 ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta 1921 aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa 1981 cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt 2041 tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt 2101 cgatcaaacc acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa 2161 aaaaaggatc tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc 2221 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct 2281 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg 2341 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc 2401 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 2461 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga 2521 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca 2581 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac 2641 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc 2701 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg 2761 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac 2821 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc 2881 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt 2941 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga 3001 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag 3061 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc 3121 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc 3181 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg 3241 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg 3301 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg 3361 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg 3421 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca 3481 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt 3541 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg 3601 aaattaatac gactcactat a //