LOCUS       sequence-74558-e        4975 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 4975)
  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 4975)
  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: pVRc37_2513
FEATURES             Location/Qualifiers
     source          1..4975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(460..477)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      630..716
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      808..835
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        854..945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1115..1774
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1181..1200)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1916..2371
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2003..2022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3057..3074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3191..3736)
                     /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(3271..3290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3808..3827
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4958..4969
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgagcagcg caccggcaac caaagaagaa gttgattttg cactgaaagt tcgtcgtgca
       61 ctggatgaac gtgcagcaag cctgccggat gcaaccaccg atcgtctggc agcagcacgt
      121 cgtgcagcac tggcacgtaa aaaaccggat gcagcaattg ttctggttcc ggcactggca
      181 ggtagcgcag gcaccctgga actgcgtccg cctggtgaac cgcgtaaaag cctggcacgt
      241 cgtctggcac gtgcatggcc tctggcactg ctgctggcag gtctgattgg tattgcatat
      301 tgggaagata tgcagcgtac cgcagaactg gcagatattg atgcagcaat gctgagcgat
      361 aatctgccgc tgaccgcata tctggatcat ggttttaatg catatctgag ccatacccat
      421 taaggatccc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
      481 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
      541 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
      601 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
      661 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
      721 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
      781 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
      841 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      901 caataaccct gataaatgct tcaataatat tgaaaaagga agactatgag tattcaacat
      961 ttccgtgtct gcttacataa acagtaatac aaggggtgtt tactagaggt tgatcgggca
     1021 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
     1081 gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca cgggatatac
     1141 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     1201 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     1261 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaacgc
     1321 tcacccggag tttcgtatgg ccatgaaaga cggtgagctg gtgatctggg atagtgttca
     1381 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcgtccctct ggagtgaata
     1441 ccacgacgat ttccggcagt ttctccacat atattcgcaa gatgtggcgt gttacggtga
     1501 aaacctggcc tatttcccta aagggtttat tgagaatatg ttttttgtct cagccaatcc
     1561 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     1621 cgttttcacg atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     1681 ccaggttcat catgccgttt gtgatggctt ccatgtcggc cgcatgctta atgaattaca
     1741 acagtactgt gatgagtggc agggcggggc gtaataatac tagctccggc aaaaaaacgg
     1801 gcaaggtgtc accaccctgc cctttttctt taaaaccgaa aagattactt cgcgtttgcc
     1861 acctgacgtc taagctgtca gaccaagttt actcatatat actttagatt gatttacgcg
     1921 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1981 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2041 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2101 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     2161 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2221 ttgttccaaa cttgaacaac actcaaccct atctcgggct attcttttga tttataaggg
     2281 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2341 aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga tcctttttga
     2401 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2461 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2521 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2581 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2641 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2701 aatcctgtta ccagtcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat
     2761 aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg
     2821 accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg
     2881 cgtagcacca ggcgtttaag ggcaccaata actgccttaa aaaaagctag cggagtgtat
     2941 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     3001 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     3061 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     3121 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     3181 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     3241 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     3301 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     3361 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     3421 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     3481 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     3541 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     3601 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     3661 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     3721 accaaaacga tctcaagaag atcatcttat taatcagata aaatatttgc tcatgagccc
     3781 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     3841 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt
     3901 gacagcttat catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa
     3961 ccctatgcta ctccctcgag ccgtcaattg tctgattcgt taccaattat taatttttaa
     4021 agtatgggca atcaattgct cctgttaaaa ttgctttaga aatactttgg cagcggtttg
     4081 ttgtattgag tttcatttgc gcattggtta aatggaaagt gacagtacgc tcactgcaac
     4141 ctaatatttt tgaaatatcc caagagcttt ttccttcgca tgcccacgct aaacattctt
     4201 tttctctttt ggttaaatcg ttgtttgatt tattatttgc tatatttatt tttcgataat
     4261 tatcaactag agaaggaaca attaatggta tgttcataca cgcatgtaaa aataaactat
     4321 ctatatagtt gtctttttct gaatgtgcaa aactaagcat tccgaagcca ttgttagccg
     4381 tatgaatagg gaaactaaac ccagtgataa gacctgatgt tttcgcttct ttaattacat
     4441 ttggagattt tttatttaca gcattgtttt caaatatatt ccaattaatt ggtgaatgat
     4501 tggagttaga ataatctact ataggatcat attttattaa attagcgtca tcataatatt
     4561 gcctccattt tttagggtaa ttatctagga ttgaaatatc agatttaacc atagaatgag
     4621 gataaatgat cgcgagtaaa taatattcac aatgtaccat tttagtcata tcagataagc
     4681 attgattaat atcattattg cttctacaag ctttaatttt attaattatt ctgtatgtgt
     4741 cgtcggcatt tatgtttttc atattcacca ccctgaattg actctcttcc gggcgctatc
     4801 atgccatacc gcgaaaggtt ttgcgccatt cgatggcgcg ccgcttcgtc aggccacata
     4861 gctttcttgt tctgatcgga acgatcgttg gctgacctgt aggatcgtac aggtttacgc
     4921 aagaaaatgg tttgttatag tcgaataaat actagagaaa gaggagaaat accat
//
