LOCUS       sequence-74557-e        5317 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 5317)
  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 5317)
  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: pVRc35_3582
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(802..819)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      972..1058
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1150..1177
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1196..1287
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1457..2116
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1523..1542)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2258..2713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2345..2364)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3399..3416)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3533..4078)
                     /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(3613..3632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4150..4169
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5300..5311
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaacccgc agaaacatag cgcaagcgtt gccgaagaac aaatgctggc acattttcgt
       61 gcacatgcac cgcagcagcc tgcaccggca ctggatcagg caattctggc agcagcacgt
      121 cgtcaggcag cacatgttga accggcacgt agttggtggc gtcgttggct ggaagcaagc
      181 cgtcgtccgc gttggcaggc agcatttgca agcctgctgg gtgttgcact ggttctgggt
      241 ctggttagcc ataatgttct ggatgatgca gaacgtcagg cacgtccgga agttgcattt
      301 agtgatgttc cgctgcgtga tggtgttgcc ggtgcagcag cagcaaaacg tgcaatgcgt
      361 gctccggcag caccggcacc gctgagcggt gaaatgagcg aaccgcctgc actgctggca
      421 agctatgcaa gcagcggtga agcaccgcag ctgatggcag aagcagcacc tccggcaccg
      481 gcagcactgg cagatcgtcc tccagcacag gcagcacagc aggcaaaagt tcaggcagcc
      541 ctggcaggcg attttgttgc acaggcacgt ggtaaagcag ttgcagttaa accggaagtt
      601 ctggatgaag cactgggtgc agttctggca ctgcgtgaac agggtaaaac cgaacaggca
      661 gcaacccagc tggcagaact gcagaaacgt tttccgggtg aaaatctggt tgaacgtctg
      721 gaacgcctgg caaccattgc agcaagcgca cgtaaacgtc cgtaaggatc cctgttttgg
      781 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
      841 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
      901 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
      961 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1021 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     1081 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     1141 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     1201 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1261 cttcaataat attgaaaaag gaagactatg agtattcaac atttccgtgt ctgcttacat
     1321 aaacagtaat acaaggggtg tttactagag gttgatcggg cacgtaagag gttccaactt
     1381 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     1441 agctaaggaa gctaaaatgg agaaaaaaat cacgggatat accaccgttg atatatccca
     1501 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     1561 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     1621 ttatccggcc tttattcaca ttcttgcccg cctgatgaac gctcacccgg agtttcgtat
     1681 ggccatgaaa gacggtgagc tggtgatctg ggatagtgtt cacccttgtt acaccgtttt
     1741 ccatgagcaa actgaaacgt tttcgtccct ctggagtgaa taccacgacg atttccggca
     1801 gtttctccac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     1861 taaagggttt attgagaata tgttttttgt ctcagccaat ccctgggtga gtttcaccag
     1921 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca cgatgggcaa
     1981 atattatacg caaggcgaca aggtgctgat gccgctggcg atccaggttc atcatgccgt
     2041 ttgtgatggc ttccatgtcg gccgcatgct taatgaatta caacagtact gtgatgagtg
     2101 gcagggcggg gcgtaataat actagctccg gcaaaaaaac gggcaaggtg tcaccaccct
     2161 gccctttttc tttaaaaccg aaaagattac ttcgcgtttg ccacctgacg tctaagctgt
     2221 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
     2281 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2341 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2401 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2461 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2521 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aacttgaaca
     2581 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     2641 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2701 acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2761 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2821 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2881 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2941 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3001 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtcag
     3061 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     3121 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt
     3181 tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcac caggcgttta
     3241 agggcaccaa taactgcctt aaaaaaagct agcggagtgt atactggctt actatgttgg
     3301 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     3361 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     3421 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     3481 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     3541 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     3601 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     3661 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     3721 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     3781 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     3841 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     3901 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     3961 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     4021 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     4081 agatcatctt attaatcaga taaaatattt gctcatgagc ccgaagtggc gagcccgatc
     4141 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     4201 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg
     4261 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccctcg
     4321 agccgtcaat tgtctgattc gttaccaatt attaattttt aaagtatggg caatcaattg
     4381 ctcctgttaa aattgcttta gaaatacttt ggcagcggtt tgttgtattg agtttcattt
     4441 gcgcattggt taaatggaaa gtgacagtac gctcactgca acctaatatt tttgaaatat
     4501 cccaagagct ttttccttcg catgcccacg ctaaacattc tttttctctt ttggttaaat
     4561 cgttgtttga tttattattt gctatattta tttttcgata attatcaact agagaaggaa
     4621 caattaatgg tatgttcata cacgcatgta aaaataaact atctatatag ttgtcttttt
     4681 ctgaatgtgc aaaactaagc attccgaagc cattgttagc cgtatgaata gggaaactaa
     4741 acccagtgat aagacctgat gttttcgctt ctttaattac atttggagat tttttattta
     4801 cagcattgtt ttcaaatata ttccaattaa ttggtgaatg attggagtta gaataatcta
     4861 ctataggatc atattttatt aaattagcgt catcataata ttgcctccat tttttagggt
     4921 aattatctag gattgaaata tcagatttaa ccatagaatg aggataaatg atcgcgagta
     4981 aataatattc acaatgtacc attttagtca tatcagataa gcattgatta atatcattat
     5041 tgcttctaca agctttaatt ttattaatta ttctgtatgt gtcgtcggca tttatgtttt
     5101 tcatattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     5161 ttttgcgcca ttcgatggcg cgccgcttcg tcaggccaca tagctttctt gttctgatcg
     5221 gaacgatcgt tggctgacct gtaggatcgt acaggtttac gcaagaaaat ggtttgttat
     5281 agtcgaataa atactagaga aagaggagaa ataccat
//
