LOCUS       sequence-74550-e        5128 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 5128)
  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 5128)
  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: pVRc25_1645
FEATURES             Location/Qualifiers
     source          1..5128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(613..630)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      783..869
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      961..988
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1007..1098
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1268..1927
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1334..1353)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2069..2524
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2156..2175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3210..3227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3344..3889)
                     /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(3424..3443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3961..3980
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5111..5122
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaacgaaa gctttagcca gcgtcagacc gatggtgatc gtcgtcgtgg tggtgatctg
       61 gaaatgaaaa aaaaccatac cagccatcag catcataccg atagcgaaac cccgatgggt
      121 cataattttg aacagtttca gcgtctgagc gcctattttg atggtgaagc aacaccggca
      181 gaacgtaaag aaattcagca tctgctggat accgatccgc aggttaaaca gcagtatcag
      241 cagctgcgtc agctgaaaca ggcactgcag ctgctgccga ttccgaccag cattagcgca
      301 cagtatctgg gtcagcgtgt tctggcacgt ctgcgtcgta gccagctgcg taccctgagc
      361 ctgtggggta gcggtgcaat tgcagcactg tttgttgccg gtgttatggg tcagatgcct
      421 cgtctgaatt ttgatcgttt tgcaaaaaac gatccggatc agcagccgac cgcagcactg
      481 gttgaaacca gtccgcaggg tgaagaagca ctggttgttg cactgaatcg tccggttctg
      541 cagattccga aactggcaac caccgaaagc ccgtaaggat ccctgttttg gcggatgaga
      601 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
      661 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
      721 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
      781 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      841 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
      901 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
      961 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     1021 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1081 tattgaaaaa ggaagactat gagtattcaa catttccgtg tctgcttaca taaacagtaa
     1141 tacaaggggt gtttactaga ggttgatcgg gcacgtaaga ggttccaact ttcaccataa
     1201 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     1261 agctaaaatg gagaaaaaaa tcacgggata taccaccgtt gatatatccc aatggcatcg
     1321 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1381 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1441 ctttattcac attcttgccc gcctgatgaa cgctcacccg gagtttcgta tggccatgaa
     1501 agacggtgag ctggtgatct gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1561 aactgaaacg ttttcgtccc tctggagtga ataccacgac gatttccggc agtttctcca
     1621 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1681 tattgagaat atgttttttg tctcagccaa tccctgggtg agtttcacca gttttgattt
     1741 aaacgtggcc aatatggaca acttcttcgc ccccgttttc acgatgggca aatattatac
     1801 gcaaggcgac aaggtgctga tgccgctggc gatccaggtt catcatgccg tttgtgatgg
     1861 cttccatgtc ggccgcatgc ttaatgaatt acaacagtac tgtgatgagt ggcagggcgg
     1921 ggcgtaataa tactagctcc ggcaaaaaaa cgggcaaggt gtcaccaccc tgcccttttt
     1981 ctttaaaacc gaaaagatta cttcgcgttt gccacctgac gtctaagctg tcagaccaag
     2041 tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg
     2101 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2161 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2221 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2281 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2341 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaacttgaac aacactcaac
     2401 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2461 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     2521 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2581 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2641 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2701 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2761 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2821 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtca ggcatttgag
     2881 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     2941 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc
     3001 tgccattcat ccgcttatta tcacttattc aggcgtagca ccaggcgttt aagggcacca
     3061 ataactgcct taaaaaaagc tagcggagtg tatactggct tactatgttg gcactgatga
     3121 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
     3181 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
     3241 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
     3301 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
     3361 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
     3421 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
     3481 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
     3541 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
     3601 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
     3661 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
     3721 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt
     3781 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
     3841 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     3901 tattaatcag ataaaatatt tgctcatgag cccgaagtgg cgagcccgat cttccccatc
     3961 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     4021 gatgcgtccg gcgtagagga tctgctcatg tttgacagct tatcatcgat gcataatgtg
     4081 cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccctc gagccgtcaa
     4141 ttgtctgatt cgttaccaat tattaatttt taaagtatgg gcaatcaatt gctcctgtta
     4201 aaattgcttt agaaatactt tggcagcggt ttgttgtatt gagtttcatt tgcgcattgg
     4261 ttaaatggaa agtgacagta cgctcactgc aacctaatat ttttgaaata tcccaagagc
     4321 tttttccttc gcatgcccac gctaaacatt ctttttctct tttggttaaa tcgttgtttg
     4381 atttattatt tgctatattt atttttcgat aattatcaac tagagaagga acaattaatg
     4441 gtatgttcat acacgcatgt aaaaataaac tatctatata gttgtctttt tctgaatgtg
     4501 caaaactaag cattccgaag ccattgttag ccgtatgaat agggaaacta aacccagtga
     4561 taagacctga tgttttcgct tctttaatta catttggaga ttttttattt acagcattgt
     4621 tttcaaatat attccaatta attggtgaat gattggagtt agaataatct actataggat
     4681 catattttat taaattagcg tcatcataat attgcctcca ttttttaggg taattatcta
     4741 ggattgaaat atcagattta accatagaat gaggataaat gatcgcgagt aaataatatt
     4801 cacaatgtac cattttagtc atatcagata agcattgatt aatatcatta ttgcttctac
     4861 aagctttaat tttattaatt attctgtatg tgtcgtcggc atttatgttt ttcatattca
     4921 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     4981 attcgatggc gcgccgcttc gtcaggccac atagctttct tgttctgatc ggaacgatcg
     5041 ttggctgacc tgtaggatcg tacaggttta cgcaagaaaa tggtttgtta tagtcgaata
     5101 aatactagag aaagaggaga aataccat
//
