LOCUS       sequence-74554-e        4873 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 4873)
  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 4873)
  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: pVRc31_34
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(358..375)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      528..614
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      706..733
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        752..843
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1013..1672
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1079..1098)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1814..2269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1901..1920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2955..2972)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3089..3634)
                     /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(3169..3188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3706..3725
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4856..4867
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaacaaag aaaaactgag cgaccacctg aaaagcgagt ggaaaaaaat cgatcagacc
       61 gcaaatccga gcattccgaa tcagaaagaa ctgctgcatc agctgagcca gatgaaagca
      121 gaatatcgta aaaaactgct gcaagaaatc attctgtttg tgttttgtgc cctgatggtt
      181 gttagcgcag caattctggc atttacccag gcaccggcag tttttattgt tctgcaggtt
      241 tgtgttctgg cagttctgcc gattctgatt gcagcagaaa aaaaacgtca tctgggtgaa
      301 tgtgaagtga aacgtggtta aggatccctg ttttggcgga tgagagaaga ttttcagcct
      361 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
      421 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
      481 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
      541 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
      601 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
      661 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
      721 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
      781 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      841 actatgagta ttcaacattt ccgtgtctgc ttacataaac agtaatacaa ggggtgttta
      901 ctagaggttg atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact
      961 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
     1021 aaaaatcacg ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     1081 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     1141 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     1201 tgcccgcctg atgaacgctc acccggagtt tcgtatggcc atgaaagacg gtgagctggt
     1261 gatctgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     1321 gtccctctgg agtgaatacc acgacgattt ccggcagttt ctccacatat attcgcaaga
     1381 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     1441 ttttgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     1501 ggacaacttc ttcgcccccg ttttcacgat gggcaaatat tatacgcaag gcgacaaggt
     1561 gctgatgccg ctggcgatcc aggttcatca tgccgtttgt gatggcttcc atgtcggccg
     1621 catgcttaat gaattacaac agtactgtga tgagtggcag ggcggggcgt aataatacta
     1681 gctccggcaa aaaaacgggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa
     1741 gattacttcg cgtttgccac ctgacgtcta agctgtcaga ccaagtttac tcatatatac
     1801 tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1861 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1921 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1981 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     2041 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2101 ttctttaata gtggactctt gttccaaact tgaacaacac tcaaccctat ctcgggctat
     2161 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2221 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta
     2281 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2341 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2401 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2461 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2521 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2581 tacatacctc gctctgctaa tcctgttacc agtcaggcat ttgagaagca cacggtcaca
     2641 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     2701 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     2761 tattatcact tattcaggcg tagcaccagg cgtttaaggg caccaataac tgccttaaaa
     2821 aaagctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt
     2881 gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg
     2941 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     3001 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     3061 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     3121 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     3181 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     3241 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     3301 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     3361 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     3421 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     3481 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     3541 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     3601 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa
     3661 atatttgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     3721 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     3781 gaggatctgc tcatgtttga cagcttatca tcgatgcata atgtgcctgt caaatggacg
     3841 aagcagggat tctgcaaacc ctatgctact ccctcgagcc gtcaattgtc tgattcgtta
     3901 ccaattatta atttttaaag tatgggcaat caattgctcc tgttaaaatt gctttagaaa
     3961 tactttggca gcggtttgtt gtattgagtt tcatttgcgc attggttaaa tggaaagtga
     4021 cagtacgctc actgcaacct aatatttttg aaatatccca agagcttttt ccttcgcatg
     4081 cccacgctaa acattctttt tctcttttgg ttaaatcgtt gtttgattta ttatttgcta
     4141 tatttatttt tcgataatta tcaactagag aaggaacaat taatggtatg ttcatacacg
     4201 catgtaaaaa taaactatct atatagttgt ctttttctga atgtgcaaaa ctaagcattc
     4261 cgaagccatt gttagccgta tgaataggga aactaaaccc agtgataaga cctgatgttt
     4321 tcgcttcttt aattacattt ggagattttt tatttacagc attgttttca aatatattcc
     4381 aattaattgg tgaatgattg gagttagaat aatctactat aggatcatat tttattaaat
     4441 tagcgtcatc ataatattgc ctccattttt tagggtaatt atctaggatt gaaatatcag
     4501 atttaaccat agaatgagga taaatgatcg cgagtaaata atattcacaa tgtaccattt
     4561 tagtcatatc agataagcat tgattaatat cattattgct tctacaagct ttaattttat
     4621 taattattct gtatgtgtcg tcggcattta tgtttttcat attcaccacc ctgaattgac
     4681 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggcgcgcc
     4741 gcttcgtcag gccacatagc tttcttgttc tgatcggaac gatcgttggc tgacctgtag
     4801 gatcgtacag gtttacgcaa gaaaatggtt tgttatagtc gaataaatac tagagaaaga
     4861 ggagaaatac cat
//
