LOCUS Exported 5194 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pVRc16_973 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5194) AUTHORS Rhodius VA, Segall-Shapiro TH, Sharon BD, Ghodasara A, Orlova E, Tabakh H, Burkhardt DH, Clancy K, Peterson TC, Gross CA, Voigt CA TITLE Design of orthogonal genetic switches based on a crosstalk map of sigmas, anti-sigmas, and promoters. JOURNAL Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58. PUBMED 24169405 REFERENCE 2 (bases 1 to 5194) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5194 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(679..696) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 849..935 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1027..1054 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1073..1164 /gene="bla" /label=AmpR promoter CDS 1334..1993 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1400..1419) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" rep_origin 2135..2590 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2222..2241) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind complement(3276..3293) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3410..3955) /direction=LEFT /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 complement(3490..3509) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4027..4046 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" RBS 5177..5188 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" ORIGIN 1 atgaaaaccg atgatctgat tgcactgctg gcagccggtg aaggtccggt tccgcgtcat 61 gcagttggtc gtcgtatggc agttgcagca ctgggtggtc tgaccgcagc actgctgctg 121 accattaccc tgtatggtgt tcgtagcgat attaccgaag ttgcacagac accgctgttt 181 tggggtaaag ttgcatttcc gaccagcctg gcactgattg gtctgtggct gaccagccgt 241 ctggcacgtc cgggtggtaa aggtgcagca ggttggaaaa tgctgggtct gccgctgctg 301 ctggtttggt gtggtgcagc agttagcatt gcgggtgcac cggttgatgc acgtgcagat 361 ctgctgtttg gtcgtacctg gcgtacctgt gcactgaata ttgccctgct gagcgttccg 421 gcatttgtta ccgttttttg ggcactgaaa ggtctggcac cgacccgtct gcgtctggct 481 ggcgcagccg gtggtctgct ggcaggtagc agcgcaaccg ttgcatattg tctgcattgt 541 ccggaaatgg gtgttccgtt ttggggtgtt tggtatgttc tgggtatgct ggttccgacc 601 gttctgggtg catggtgggg tccgcgtatg ctgcgttggt aaggatccct gttttggcgg 661 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa 721 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga 781 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg 841 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt 901 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg 961 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa 1021 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta 1081 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 1141 caataatatt gaaaaaggaa gactatgagt attcaacatt tccgtgtctg cttacataaa 1201 cagtaataca aggggtgttt actagaggtt gatcgggcac gtaagaggtt ccaactttca 1261 ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc 1321 taaggaagct aaaatggaga aaaaaatcac gggatatacc accgttgata tatcccaatg 1381 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac 1441 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta 1501 tccggccttt attcacattc ttgcccgcct gatgaacgct cacccggagt ttcgtatggc 1561 catgaaagac ggtgagctgg tgatctggga tagtgttcac ccttgttaca ccgttttcca 1621 tgagcaaact gaaacgtttt cgtccctctg gagtgaatac cacgacgatt tccggcagtt 1681 tctccacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa 1741 agggtttatt gagaatatgt tttttgtctc agccaatccc tgggtgagtt tcaccagttt 1801 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacga tgggcaaata 1861 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatc caggttcatc atgccgtttg 1921 tgatggcttc catgtcggcc gcatgcttaa tgaattacaa cagtactgtg atgagtggca 1981 gggcggggcg taataatact agctccggca aaaaaacggg caaggtgtca ccaccctgcc 2041 ctttttcttt aaaaccgaaa agattacttc gcgtttgcca cctgacgtct aagctgtcag 2101 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc 2161 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 2221 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 2281 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 2341 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 2401 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac ttgaacaaca 2461 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 2521 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg 2581 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 2641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 2701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 2761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 2821 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 2881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtcaggca 2941 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag 3001 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg 3061 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag gcgtttaagg 3121 gcaccaataa ctgccttaaa aaaagctagc ggagtgtata ctggcttact atgttggcac 3181 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt 3241 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg 3301 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat 3361 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc 3421 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga 3481 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc 3541 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc 3601 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc 3661 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga 3721 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa 3781 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag 3841 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa 3901 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga 3961 tcatcttatt aatcagataa aatatttgct catgagcccg aagtggcgag cccgatcttc 4021 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc 4081 ggccacgatg cgtccggcgt agaggatctg ctcatgtttg acagcttatc atcgatgcat 4141 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccctcgagc 4201 cgtcaattgt ctgattcgtt accaattatt aatttttaaa gtatgggcaa tcaattgctc 4261 ctgttaaaat tgctttagaa atactttggc agcggtttgt tgtattgagt ttcatttgcg 4321 cattggttaa atggaaagtg acagtacgct cactgcaacc taatattttt gaaatatccc 4381 aagagctttt tccttcgcat gcccacgcta aacattcttt ttctcttttg gttaaatcgt 4441 tgtttgattt attatttgct atatttattt ttcgataatt atcaactaga gaaggaacaa 4501 ttaatggtat gttcatacac gcatgtaaaa ataaactatc tatatagttg tctttttctg 4561 aatgtgcaaa actaagcatt ccgaagccat tgttagccgt atgaataggg aaactaaacc 4621 cagtgataag acctgatgtt ttcgcttctt taattacatt tggagatttt ttatttacag 4681 cattgttttc aaatatattc caattaattg gtgaatgatt ggagttagaa taatctacta 4741 taggatcata ttttattaaa ttagcgtcat cataatattg cctccatttt ttagggtaat 4801 tatctaggat tgaaatatca gatttaacca tagaatgagg ataaatgatc gcgagtaaat 4861 aatattcaca atgtaccatt ttagtcatat cagataagca ttgattaata tcattattgc 4921 ttctacaagc tttaatttta ttaattattc tgtatgtgtc gtcggcattt atgtttttca 4981 tattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt 5041 tgcgccattc gatggcgcgc cgcttcgtca ggccacatag ctttcttgtt ctgatcggaa 5101 cgatcgttgg ctgacctgta ggatcgtaca ggtttacgca agaaaatggt ttgttatagt 5161 cgaataaata ctagagaaag aggagaaata ccat //