LOCUS       sequence-74543-e        4759 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 4759)
  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 4759)
  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: pVRc15_524
FEATURES             Location/Qualifiers
     source          1..4759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(244..261)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      414..500
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      592..619
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        638..729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             899..1558
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(965..984)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1700..2155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1787..1806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2841..2858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2975..3520)
                     /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(3055..3074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3592..3611
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4742..4753
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaactttg gcgtcgaaga tatgattgaa catgttccga tggaagataa acgtaaaggt
       61 gcagcagcac tggatgaagc acgtctgcgt cagcaggcaa ttggtgttaa actgcgtcag
      121 atgtttgatg aagttgttaa tgaaccggtg ccggatgaat ttctggcaat tctgcgtaaa
      181 gcagaacgtc cggcaggcgg tgaataagga tccctgtttt ggcggatgag agaagatttt
      241 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
      301 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
      361 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
      421 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
      481 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
      541 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
      601 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
      661 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      721 aggaagacta tgagtattca acatttccgt gtctgcttac ataaacagta atacaagggg
      781 tgtttactag aggttgatcg ggcacgtaag aggttccaac tttcaccata atgaaataag
      841 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
      901 ggagaaaaaa atcacgggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
      961 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     1021 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     1081 cattcttgcc cgcctgatga acgctcaccc ggagtttcgt atggccatga aagacggtga
     1141 gctggtgatc tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     1201 gttttcgtcc ctctggagtg aataccacga cgatttccgg cagtttctcc acatatattc
     1261 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     1321 tatgtttttt gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     1381 caatatggac aacttcttcg cccccgtttt cacgatgggc aaatattata cgcaaggcga
     1441 caaggtgctg atgccgctgg cgatccaggt tcatcatgcc gtttgtgatg gcttccatgt
     1501 cggccgcatg cttaatgaat tacaacagta ctgtgatgag tggcagggcg gggcgtaata
     1561 atactagctc cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac
     1621 cgaaaagatt acttcgcgtt tgccacctga cgtctaagct gtcagaccaa gtttactcat
     1681 atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1741 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1801 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1861 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     1921 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1981 tccacgttct ttaatagtgg actcttgttc caaacttgaa caacactcaa ccctatctcg
     2041 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2101 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaaag
     2161 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2221 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2281 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2341 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2401 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2461 accgcctaca tacctcgctc tgctaatcct gttaccagtc aggcatttga gaagcacacg
     2521 gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt
     2581 taacgaccct gccctgaacc gacgaccggg tcgaatttgc tttcgaattt ctgccattca
     2641 tccgcttatt atcacttatt caggcgtagc accaggcgtt taagggcacc aataactgcc
     2701 ttaaaaaaag ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     2761 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     2821 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     2881 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     2941 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     3001 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     3061 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     3121 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     3181 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     3241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     3301 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     3361 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     3421 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     3481 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     3541 gataaaatat ttgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     3601 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     3661 ggcgtagagg atctgctcat gtttgacagc ttatcatcga tgcataatgt gcctgtcaaa
     3721 tggacgaagc agggattctg caaaccctat gctactccct cgagccgtca attgtctgat
     3781 tcgttaccaa ttattaattt ttaaagtatg ggcaatcaat tgctcctgtt aaaattgctt
     3841 tagaaatact ttggcagcgg tttgttgtat tgagtttcat ttgcgcattg gttaaatgga
     3901 aagtgacagt acgctcactg caacctaata tttttgaaat atcccaagag ctttttcctt
     3961 cgcatgccca cgctaaacat tctttttctc ttttggttaa atcgttgttt gatttattat
     4021 ttgctatatt tatttttcga taattatcaa ctagagaagg aacaattaat ggtatgttca
     4081 tacacgcatg taaaaataaa ctatctatat agttgtcttt ttctgaatgt gcaaaactaa
     4141 gcattccgaa gccattgtta gccgtatgaa tagggaaact aaacccagtg ataagacctg
     4201 atgttttcgc ttctttaatt acatttggag attttttatt tacagcattg ttttcaaata
     4261 tattccaatt aattggtgaa tgattggagt tagaataatc tactatagga tcatatttta
     4321 ttaaattagc gtcatcataa tattgcctcc attttttagg gtaattatct aggattgaaa
     4381 tatcagattt aaccatagaa tgaggataaa tgatcgcgag taaataatat tcacaatgta
     4441 ccattttagt catatcagat aagcattgat taatatcatt attgcttcta caagctttaa
     4501 ttttattaat tattctgtat gtgtcgtcgg catttatgtt tttcatattc accaccctga
     4561 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     4621 cgcgccgctt cgtcaggcca catagctttc ttgttctgat cggaacgatc gttggctgac
     4681 ctgtaggatc gtacaggttt acgcaagaaa atggtttgtt atagtcgaat aaatactaga
     4741 gaaagaggag aaataccat
//
