LOCUS . 3829 bp DNA circular UNK 01-JAN-1980 DEFINITION . ACCESSION VERSION KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers rep_origin complement(1662..2411) /direction=LEFT /note="Plasmids containing the RSF 1030 origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." /note="color: #ffff00" /label="RSF ori" RBS 286..291 /note="ribosome binding site" /note="color: #a6acb3" /label="RBS" RBS 58..63 /note="ribosome binding site" /note="color: #a6acb3" /label="RBS" CDS 300..302 /codon_start=1 /product="start codon" /transl_table=1 /translation="M" /label="ATG" /note="color: #993366" CDS 71..73 /codon_start=1 /product="start codon" /transl_table=1 /translation="M" /label="ATG" /note="color: #993366" misc_feature 69..168 /note="multiple cloning site 1" /note="color: #99ccff" /label="MCS-1" misc_feature 297..438 /note="multiple cloning site 2" /note="color: #99ccff" /label="MCS-2" CDS 83..100 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" /note="color: #cc99b2" promoter 2..3813 /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff" /label="T7 promoter" promoter 214..232 /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff" /label="T7 promoter" protein_bind 233..257 /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" 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" CDS 366..410 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /transl_table=1 /translation="KETAAAKFERQHMDS" /label="S-Tag" /note="color: #cc99b2" terminator 462..509 /note="transcription terminator for bacteriophage T7 RNA polymerase" /note="color: #ffffff" /label="T7 terminator" promoter complement(3690..3767) /gene="lacI" /note="" /note="color: #ffffff" /label="lacI promoter" promoter complement(1555..1646) /gene="bla" /label="AmpR promoter" /note="color: #ffffff" CDS complement(739..1554) /codon_start=1 /gene="aph(3')-Ia" /note="confers resistance to kanamycin" /note="color: #ccffcc" /product="aminoglycoside phosphotransferase" /transl_table=1 /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*" /label="KanR" CDS complement(2607..3689) /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" ORIGIN 1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag 61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag 121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag 181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt 241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata 301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt 361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg 421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg 481 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg 541 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt 601 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt 661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 721 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt 781 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa 841 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg 901 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa 961 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca 1021 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc 1081 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca 1141 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt 1201 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt 1261 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat 1321 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc 1381 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt 1441 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat 1501 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt 1561 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 1621 tgaatgtatt tagaaaaata aacaaatagg catgcagcgc tcttccgctt cctcgctcac 1681 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact caaaagcggt 1741 aatacggtta tccacagaat caggggataa agccggaaag aacatgtgag caaaaagcaa 1801 agcaccggaa gaagccaacg ccgcaggcgt ttttccatag gctccgcccc cctgacgagc 1861 atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc gacaggacta taaagatacc 1921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 1981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt 2041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 2101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 2161 acgacttatc gccactggca gcagccattg gtaactgatt tagaggactt tgtcttgaag 2221 ttatgcacct gttaaggcta aactgaaaga acagattttg gtgagtgcgg tcctccaacc 2281 cacttacctt ggttcaaaga gttggtagct cagcgaacct tgagaaaacc accgttggta 2341 gcggtggttt ttctttattt atgagatgat gaatcaatcg gtctatcaag tcaacgaaca 2401 gctattccgt tactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc 2461 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg 2521 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga 2581 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 2641 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc 2701 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg 2761 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt 2821 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact 2881 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc 2941 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc 3001 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga 3061 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa 3121 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg 3181 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag 3241 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg 3301 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc 3361 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc 3421 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga 3481 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg 3541 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa 3601 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct 3661 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg 3721 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg 3781 ctctccctta tgcgactcct gcattaggaa attaatacga ctcactata //