LOCUS       sequence-74559-e        5917 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 5917)
  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 5917)
  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: pVRc38_1322
FEATURES             Location/Qualifiers
     source          1..5917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1402..1419)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1572..1658
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1750..1777
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1796..1887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2057..2716
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2123..2142)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2858..3313
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2945..2964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3999..4016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4133..4678)
                     /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(4213..4232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4750..4769
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5900..5911
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgtccgatg gtaatgatgc acgtccggat gcacgtgatg gtcatgatcc aggtagtgat
       61 gcccgtgatg gccacgatcc tggttcacat gcgcgtgacg gccatgatcc aggtgcaggt
      121 ccgtttgatg gccatggtcc gggttcagat gctggtgatg gtgatgatgc cggtcctgac
      181 gcacgcgacg gtcatggtgc accggcaggt cctgcaggtc gtgaacagcg taaacatccg
      241 catgataccc atgatatgca cgatacacat gatacgcatg caggtaaagg caccgttaat
      301 catggtccgg cagcacaggg tcctgatgca ccgggtgcag atggtccggc aaccgatgaa
      361 gcagcactgc gtgcaatgat gcagcgtgca gttcgtgaaa tggaaccgag tgatggcacc
      421 ctggaacatc tgcgtcgtgc tgttccggca cgtcgtgcac gtaaacgtca ggcactggtt
      481 ggtgcagcag cagcagccct gtttctgggc acagccgttc cggcactggt tcatgttagc
      541 aatgcaaccg gtgccggtgc agatccgagc gttgcaggta atgcaagcca ggcacagggt
      601 ggtgcaagcc agggtaaaga tccggcaggc ggtcagagcg gtgttgcagg caccggtgat
      661 acaccggaag atcgtgataa agcagatccg aaagaaacac cgggtggtaa agaaccgggt
      721 gcagcaaccg gtgcaccgcc tagcggtgtt ccgagcgcaa gcagtccggc agatgttccg
      781 gcatgtgcac cgggtagcct gggtccggca gttgcaagca gcgcagaacc ggatagcacc
      841 ggtgttgttt atggtagctt tcgtgttacc aatgttagca gtgatggttg taccgttacc
      901 ggtccgggta cagttgttac cgcaagcctg ggtgcagcag aagcaacccg tattggcacc
      961 gcacgtcatg cagccggtga tgcagcagca ggtctgcctg atccgagcct ggaaaccgcc
     1021 agcctggcac tggcacctgg tgcagcctat gaagttcagt ttgcatgggt tccgagcgaa
     1081 acctgtccga ccaccggtgg caccacaggt ggtggtagcg gtggtccgag tccagatccg
     1141 agcccgaccg cagataccac tgcagccggt ggtacaagtg ccggtggtgg tgaagcaggt
     1201 ccgaccaccc agctgattac cgaagatggt cctgcagaag gtagcgttag cgttacctat
     1261 actccggaag gtggtagtgg tagcgcaacc gcaaccgtta gcaatgcctg tgcaggcacc
     1321 gtttattgga ccggtctgct ggcagatagc ggtagcggtg cataaggatc cctgttttgg
     1381 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     1441 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     1501 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     1561 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1621 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     1681 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     1741 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     1801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1861 cttcaataat attgaaaaag gaagactatg agtattcaac atttccgtgt ctgcttacat
     1921 aaacagtaat acaaggggtg tttactagag gttgatcggg cacgtaagag gttccaactt
     1981 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     2041 agctaaggaa gctaaaatgg agaaaaaaat cacgggatat accaccgttg atatatccca
     2101 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     2161 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     2221 ttatccggcc tttattcaca ttcttgcccg cctgatgaac gctcacccgg agtttcgtat
     2281 ggccatgaaa gacggtgagc tggtgatctg ggatagtgtt cacccttgtt acaccgtttt
     2341 ccatgagcaa actgaaacgt tttcgtccct ctggagtgaa taccacgacg atttccggca
     2401 gtttctccac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     2461 taaagggttt attgagaata tgttttttgt ctcagccaat ccctgggtga gtttcaccag
     2521 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca cgatgggcaa
     2581 atattatacg caaggcgaca aggtgctgat gccgctggcg atccaggttc atcatgccgt
     2641 ttgtgatggc ttccatgtcg gccgcatgct taatgaatta caacagtact gtgatgagtg
     2701 gcagggcggg gcgtaataat actagctccg gcaaaaaaac gggcaaggtg tcaccaccct
     2761 gccctttttc tttaaaaccg aaaagattac ttcgcgtttg ccacctgacg tctaagctgt
     2821 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
     2881 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2941 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3001 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3061 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3121 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aacttgaaca
     3181 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     3241 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3301 acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3361 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3421 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3481 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3541 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3601 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtcag
     3661 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     3721 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt
     3781 tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcac caggcgttta
     3841 agggcaccaa taactgcctt aaaaaaagct agcggagtgt atactggctt actatgttgg
     3901 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     3961 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     4021 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     4081 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     4141 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     4201 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     4261 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     4321 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     4381 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     4441 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     4501 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     4561 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     4621 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     4681 agatcatctt attaatcaga taaaatattt gctcatgagc ccgaagtggc gagcccgatc
     4741 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     4801 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg
     4861 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccctcg
     4921 agccgtcaat tgtctgattc gttaccaatt attaattttt aaagtatggg caatcaattg
     4981 ctcctgttaa aattgcttta gaaatacttt ggcagcggtt tgttgtattg agtttcattt
     5041 gcgcattggt taaatggaaa gtgacagtac gctcactgca acctaatatt tttgaaatat
     5101 cccaagagct ttttccttcg catgcccacg ctaaacattc tttttctctt ttggttaaat
     5161 cgttgtttga tttattattt gctatattta tttttcgata attatcaact agagaaggaa
     5221 caattaatgg tatgttcata cacgcatgta aaaataaact atctatatag ttgtcttttt
     5281 ctgaatgtgc aaaactaagc attccgaagc cattgttagc cgtatgaata gggaaactaa
     5341 acccagtgat aagacctgat gttttcgctt ctttaattac atttggagat tttttattta
     5401 cagcattgtt ttcaaatata ttccaattaa ttggtgaatg attggagtta gaataatcta
     5461 ctataggatc atattttatt aaattagcgt catcataata ttgcctccat tttttagggt
     5521 aattatctag gattgaaata tcagatttaa ccatagaatg aggataaatg atcgcgagta
     5581 aataatattc acaatgtacc attttagtca tatcagataa gcattgatta atatcattat
     5641 tgcttctaca agctttaatt ttattaatta ttctgtatgt gtcgtcggca tttatgtttt
     5701 tcatattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     5761 ttttgcgcca ttcgatggcg cgccgcttcg tcaggccaca tagctttctt gttctgatcg
     5821 gaacgatcgt tggctgacct gtaggatcgt acaggtttac gcaagaaaat ggtttgttat
     5881 agtcgaataa atactagaga aagaggagaa ataccat
//
