LOCUS Exported 5222 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PTac T3 sigma fragment expression. ACCESSION . VERSION . KEYWORDS pTHSSd_24 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5222) AUTHORS Segall-Shapiro TH, Meyer AJ, Ellington AD, Sontag ED, Voigt CA TITLE A 'resource allocator' for transcription based on a highly fragmented T7 RNA polymerase. JOURNAL Mol Syst Biol. 2014 Jul 30;10(7):742. PUBMED 25080493 REFERENCE 2 (bases 1 to 5222) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5222 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 1040..1071 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" rep_origin 1582..2126 /direction=RIGHT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind 2027..2046 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2243..2260 /label=L4440 /note="L4440 vector, forward primer" CDS 2835..3626 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" terminator 3753..3782 /label=T3Te terminator /note="phage T3 early transcription terminator" CDS complement(3845..4927) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 4889..4908 /label=LacI-R /note="LacI, reverse primer" promoter 5062..5090 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 5096..5115 /label=lac operator (symmetric) /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-beta-D-thiogalactopyranoside (IPTG). The symmetric lac operator was optimized for tight binding of lac repressor." misc_RNA 5131..5180 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" ORIGIN 1 atgagcatcg cggcgaccct ggagaacgat ctggcgcgtc tggaaaacga aaacgctcgt 61 ctcgaaaaag acatcgcgaa cctggaacgt gacctggcga aactggagcg tgaagaagcg 121 tacttcggag gttcaggtgg taagaacact ggtgaaatct ctgagaaagt caagctgggc 181 actaaggcac tggctggtca atggctggct tacggtgtta ctcgcagtgt gactaagcgt 241 tcagtcatga cgctggctta cgggtccaaa gagttcggct tccgtcaaca agtgctggaa 301 gataccattc agccagctat tgattccggc aagggtctga tgttcactca gccgaatcag 361 gctgctggat acatggctaa gctgatttgg gaatctgtga gcgtgacggt ggtagctgcg 421 gttgaagcaa tgaactggct taagtctgct gctaagctgc tggctgctga ggtcaaagat 481 aagaagactg gagagattct tcgcaagcgt tgcgctgtgc attgggtaac tcctgatggt 541 ttccctgtgt ggcaggaata caagaagcct attcagaagc gcctggacat gattttcttg 601 ggtcaatttc gcttgcaacc taccattaac accaacaaag atagcgagat tgatgcacac 661 aaacaggagt ctggtatcgc tcctaacttt gtacacagcc aagacggtag ccaccttcgt 721 aagactgtag tgtgggcaca cgagaagtac ggaatcgaat cttttgcact gattcacgac 781 tccttcggta cgattccggc tgacgctgcg aacctgttca aagcagtgcg cgaaactatg 841 gttgacacat atgagtcttg tgatgtactg gctgatttct acgaccagtt cgctgaccag 901 ttgcacgagt ctcaattgga caaaatgcca gcacttccgg ctaaaggtaa cttgaacctc 961 cgtgacatct tagagtcgga cttcgcgttc gcgtaacgct gcaggagtca ctaagggtta 1021 gttagttaga ttagcagaaa gtcaaaagcc tccgaccgga ggcttttgac taaaacttcc 1081 cttggggtta tcattggggc tcactcaaag gcggtaatca gataaaaaaa atccttagct 1141 ttcgctaagg atgatttctg ctagagatgg aatagactgg atggaggcgg ataaagttgc 1201 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 1261 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 1321 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 1381 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 1441 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 1501 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 1561 ccccttaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga 1621 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt 1681 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc 1741 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca 1801 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc 1861 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc 1921 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg 1981 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccagggga aacgcctggt 2041 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct 2101 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt 2161 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg 2221 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta 2281 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca 2341 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgaggtc 2401 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc 2461 agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt 2521 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct 2581 tcaactcagc aaaagttcga tttattcaac aaagccacgt tgtgtctcaa aatctctgat 2641 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa 2701 acagtaatac aaggggtgtt tactagaggt tgatcgggca cgtaagaggt tccaactttc 2761 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag 2821 ctaaggaagc taaaatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg 2881 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct 2941 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg 3001 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt 3061 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca 3121 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg 3181 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga 3241 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc 3301 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc 3361 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca 3421 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc 3481 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag 3541 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg 3601 agatcaccaa ggtagtcggc aaataatact agctccggca aaaaaacggg caaggtgtca 3661 ccaccctgcc ctttttcttt aaaaccgaaa agattacttc gcgtttgcca cctgacgtct 3721 aagaaaagga atattcagca atttgcccgt gccgaagaaa ggcccacccg tgaaggtgag 3781 ccagtgagtt gattgctacg taatgtcggc caattcgcgc taacttacat taattgcgtt 3841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 3901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca 3961 gtgagactgg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc 4021 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga 4081 tataacatga gctatcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc 4141 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca 4201 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca 4261 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt 4321 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg 4381 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcctcat 4441 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa 4501 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga 4561 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc 4621 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa 4681 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca 4741 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg 4801 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca 4861 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg 4921 gtttcatatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa 4981 aggttttgcg ccattcgatg gcgcgccgct tcgtcaggcc acatagcttt cttgttctga 5041 tcggaacgat cgttggctgt gttgacaatt aatcatcggc tcgtataatg tgtggaattg 5101 tgagcgctca caattctatg gactatgttt ctgtcaccgg atgtgctttc cggtctgatg 5161 agtccgtgag gacgaaacag cctctacaaa taattttgtt taatcacaca ggaaagtact 5221 ag //