LOCUS       sequence-74535-e        5203 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 5203)
  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 5203)
  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: pVRc01_3473
FEATURES             Location/Qualifiers
     source          1..5203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(688..705)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      858..944
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1036..1063
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1082..1173
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1343..2002
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1409..1428)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2144..2599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2231..2250)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3285..3302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3419..3964)
                     /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(3499..3518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4036..4055
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5186..5197
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgagcaact atgataatgc agaactgctg gcactgtggc tggaaggtaa actgaccccg
       61 aaacagcgtg atgaatttga acagcgttgt attcaggata gccagtttgc agaacaggtt
      121 gatgcagcag caatggttaa aattggtgca gatgattata gcagcagcga agttccgcag
      181 tggaataaag cagcaacctt tgaaccgcag caggcaaata attcaggtag ctggctgagc
      241 ggtgttagcc tggttaccag cgcactggca attgttctgg ttctgaccgg cacccaggtt
      301 accaccagtg aaggtgaact gcgtattcgt tttggtagcg gtcagagcga acaggcactg
      361 gttaaactgg ttgatagcaa actggatgcc tttaaacaga atcagcagga tgcatttacc
      421 ctgtatgcac agaccctgca gcagcagcag agcgaaagcg caagccagct gaccaattat
      481 ctgctgagca gcagccgtaa agaacgtcgt gaagattttg cagaactgat caaatttgtg
      541 aatcagcagc gtagtgatga tcagctgttt tatgcacgtc agctgaatca gctgcagcag
      601 gatgtttatc atgatgcaac cggtgttgca ctggaaagca ttcagcagta aggatccctg
      661 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
      721 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
      781 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
      841 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
      901 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
      961 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     1021 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     1081 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1141 taaatgcttc aataatattg aaaaaggaag actatgagta ttcaacattt ccgtgtctgc
     1201 ttacataaac agtaatacaa ggggtgttta ctagaggttg atcgggcacg taagaggttc
     1261 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
     1321 ttcaggagct aaggaagcta aaatggagaa aaaaatcacg ggatatacca ccgttgatat
     1381 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1441 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1501 caagttttat ccggccttta ttcacattct tgcccgcctg atgaacgctc acccggagtt
     1561 tcgtatggcc atgaaagacg gtgagctggt gatctgggat agtgttcacc cttgttacac
     1621 cgttttccat gagcaaactg aaacgttttc gtccctctgg agtgaatacc acgacgattt
     1681 ccggcagttt ctccacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     1741 tttccctaaa gggtttattg agaatatgtt ttttgtctca gccaatccct gggtgagttt
     1801 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcacgat
     1861 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgatcc aggttcatca
     1921 tgccgtttgt gatggcttcc atgtcggccg catgcttaat gaattacaac agtactgtga
     1981 tgagtggcag ggcggggcgt aataatacta gctccggcaa aaaaacgggc aaggtgtcac
     2041 caccctgccc tttttcttta aaaccgaaaa gattacttcg cgtttgccac ctgacgtcta
     2101 agctgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
     2161 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2221 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2281 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2341 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2401 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2461 tgaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     2521 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2581 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2641 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2701 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2761 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2821 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2881 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2941 agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     3001 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat
     3061 ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg
     3121 cgtttaaggg caccaataac tgccttaaaa aaagctagcg gagtgtatac tggcttacta
     3181 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     3241 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     3301 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     3361 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     3421 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     3481 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
     3541 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
     3601 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     3661 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3721 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     3781 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     3841 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     3901 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     3961 tcaagaagat catcttatta atcagataaa atatttgctc atgagcccga agtggcgagc
     4021 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     4081 ggtgatgccg gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagcttatca
     4141 tcgatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
     4201 ccctcgagcc gtcaattgtc tgattcgtta ccaattatta atttttaaag tatgggcaat
     4261 caattgctcc tgttaaaatt gctttagaaa tactttggca gcggtttgtt gtattgagtt
     4321 tcatttgcgc attggttaaa tggaaagtga cagtacgctc actgcaacct aatatttttg
     4381 aaatatccca agagcttttt ccttcgcatg cccacgctaa acattctttt tctcttttgg
     4441 ttaaatcgtt gtttgattta ttatttgcta tatttatttt tcgataatta tcaactagag
     4501 aaggaacaat taatggtatg ttcatacacg catgtaaaaa taaactatct atatagttgt
     4561 ctttttctga atgtgcaaaa ctaagcattc cgaagccatt gttagccgta tgaataggga
     4621 aactaaaccc agtgataaga cctgatgttt tcgcttcttt aattacattt ggagattttt
     4681 tatttacagc attgttttca aatatattcc aattaattgg tgaatgattg gagttagaat
     4741 aatctactat aggatcatat tttattaaat tagcgtcatc ataatattgc ctccattttt
     4801 tagggtaatt atctaggatt gaaatatcag atttaaccat agaatgagga taaatgatcg
     4861 cgagtaaata atattcacaa tgtaccattt tagtcatatc agataagcat tgattaatat
     4921 cattattgct tctacaagct ttaattttat taattattct gtatgtgtcg tcggcattta
     4981 tgtttttcat attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     5041 gaaaggtttt gcgccattcg atggcgcgcc gcttcgtcag gccacatagc tttcttgttc
     5101 tgatcggaac gatcgttggc tgacctgtag gatcgtacag gtttacgcaa gaaaatggtt
     5161 tgttatagtc gaataaatac tagagaaaga ggagaaatac cat
//
