LOCUS       sequence-74536-e        5149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5149)
  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 5149)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Syst 
            Biol."; date: "2013-10-29"; volume: "9:702. doi"; pages: " 
            10.1038/msb.2013.58"
COMMENT     Sequence Label: pVRc02_915
FEATURES             Location/Qualifiers
     source          1..5149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(634..651)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      804..890
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      982..1009
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1028..1119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1289..1948
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1355..1374)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2090..2545
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2177..2196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3231..3248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3365..3910)
                     /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(3445..3464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3982..4001
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5132..5143
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atggaaaaaa ccggtcaaga atgggttagc gcaagcgttg atggtgaaac cgatcgtcag
       61 accctggcag aactggcagc agataccgat agccatgcaa aatggcagcg ttatcatctg
      121 atgggtgatg caatgcgtga tgaactgccg caggcaatgc cgctggatct gagcgcacgt
      181 attgcagcag cactggaaga tgaaccgacc attctggcac cgaaagttga acgtgaagca
      241 acccaggcac cgagccgtgc agttgttgtt ccgtttatgc gtcagctggg tcagtatggt
      301 attgcagccg cagttgcact gatggcagtt gttggtgttc agaattatca gagcacccag
      361 gatgatgcac cgctgccggt tctgaatacc cgtccgctgg ttggcaccgc aacaccggtt
      421 agcctgcaga ccggtccggt tgcaaatcag aatcagggta atgcaaatga tcagctgctg
      481 gaacagcgtc gtcgtattaa tgcatatctg caggatcaca tgctgcagca gcgtctgaat
      541 accggtgccg ttgttgatga taattcagaa gttaccccga ttccggttaa tcgttaagga
      601 tccctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
      661 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
      721 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
      781 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      841 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
      901 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
      961 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     1021 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1081 ccctgataaa tgcttcaata atattgaaaa aggaagacta tgagtattca acatttccgt
     1141 gtctgcttac ataaacagta atacaagggg tgtttactag aggttgatcg ggcacgtaag
     1201 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
     1261 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcacgggat ataccaccgt
     1321 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     1381 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     1441 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga acgctcaccc
     1501 ggagtttcgt atggccatga aagacggtga gctggtgatc tgggatagtg ttcacccttg
     1561 ttacaccgtt ttccatgagc aaactgaaac gttttcgtcc ctctggagtg aataccacga
     1621 cgatttccgg cagtttctcc acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     1681 ggcctatttc cctaaagggt ttattgagaa tatgtttttt gtctcagcca atccctgggt
     1741 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     1801 cacgatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgatccaggt
     1861 tcatcatgcc gtttgtgatg gcttccatgt cggccgcatg cttaatgaat tacaacagta
     1921 ctgtgatgag tggcagggcg gggcgtaata atactagctc cggcaaaaaa acgggcaagg
     1981 tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt tgccacctga
     2041 cgtctaagct gtcagaccaa gtttactcat atatacttta gattgattta cgcgccctgt
     2101 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2161 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2221 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2281 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2341 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2401 caaacttgaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     2461 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2521 aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt ttgataatct
     2581 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2761 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2881 gttaccagtc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     2941 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     3001 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     3061 accaggcgtt taagggcacc aataactgcc ttaaaaaaag ctagcggagt gtatactggc
     3121 ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag
     3181 gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact
     3241 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
     3301 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
     3361 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
     3421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg
     3481 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
     3541 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt
     3601 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3661 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
     3721 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga
     3781 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
     3841 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
     3901 acgatctcaa gaagatcatc ttattaatca gataaaatat ttgctcatga gcccgaagtg
     3961 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     4021 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc
     4081 ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     4141 gctactccct cgagccgtca attgtctgat tcgttaccaa ttattaattt ttaaagtatg
     4201 ggcaatcaat tgctcctgtt aaaattgctt tagaaatact ttggcagcgg tttgttgtat
     4261 tgagtttcat ttgcgcattg gttaaatgga aagtgacagt acgctcactg caacctaata
     4321 tttttgaaat atcccaagag ctttttcctt cgcatgccca cgctaaacat tctttttctc
     4381 ttttggttaa atcgttgttt gatttattat ttgctatatt tatttttcga taattatcaa
     4441 ctagagaagg aacaattaat ggtatgttca tacacgcatg taaaaataaa ctatctatat
     4501 agttgtcttt ttctgaatgt gcaaaactaa gcattccgaa gccattgtta gccgtatgaa
     4561 tagggaaact aaacccagtg ataagacctg atgttttcgc ttctttaatt acatttggag
     4621 attttttatt tacagcattg ttttcaaata tattccaatt aattggtgaa tgattggagt
     4681 tagaataatc tactatagga tcatatttta ttaaattagc gtcatcataa tattgcctcc
     4741 attttttagg gtaattatct aggattgaaa tatcagattt aaccatagaa tgaggataaa
     4801 tgatcgcgag taaataatat tcacaatgta ccattttagt catatcagat aagcattgat
     4861 taatatcatt attgcttcta caagctttaa ttttattaat tattctgtat gtgtcgtcgg
     4921 catttatgtt tttcatattc accaccctga attgactctc ttccgggcgc tatcatgcca
     4981 taccgcgaaa ggttttgcgc cattcgatgg cgcgccgctt cgtcaggcca catagctttc
     5041 ttgttctgat cggaacgatc gttggctgac ctgtaggatc gtacaggttt acgcaagaaa
     5101 atggtttgtt atagtcgaat aaatactaga gaaagaggag aaataccat
//
