LOCUS       sequence-74545-e        5197 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 5197)
  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 5197)
  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: pVRc16_3622
FEATURES             Location/Qualifiers
     source          1..5197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(682..699)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      852..938
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1030..1057
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1076..1167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1337..1996
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1403..1422)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2138..2593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2225..2244)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3279..3296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3413..3958)
                     /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(3493..3512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4030..4049
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5180..5191
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgatgaaaa ccgatgaact gattagcctg ctggcaaccg cagaaggtcc ggttgatcgt
       61 catgcactgg cacgtcgtct gggtctggca ctgctggcag gtctgctggg tgcactgctg
      121 ctgaccgttg cactgtatgg tgttcgtagc gatctggcag aagttgcacg tacaccgctg
      181 ttttgggcaa aagttgcact gccgaccagc ctggccctgc tgggtctgtg gctgacccag
      241 cgtctggcac gtccgggtgt tcgtggtggt gcactgtggg gtctgctggg cgttccgctg
      301 ctgctggttt ggctgggtgc agcaattagc ctgtttggtg caccgcctga agcacgtgca
      361 gatctgattt ttggtcgtac ctggcgtacc tgtgcactga atattaccct gctgagcaca
      421 ccggttttta ttgcagtttt ttgggcactg cgtggtctgg caccgacccg tctgcgtcag
      481 gctggtgcag ccggtggtct gctggctggt agcaccgcaa ccctggttta ttgtctgcat
      541 tgtccggaaa tgggtgttcc gttttggggt ctgtggtatc tgctgggtat gctggttccg
      601 accctgctgg gtgccgttct gggtccgcgt ctgctgcgtt ggtaaggatc cctgttttgg
      661 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
      721 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
      781 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
      841 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
      901 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
      961 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     1021 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     1081 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1141 cttcaataat attgaaaaag gaagactatg agtattcaac atttccgtgt ctgcttacat
     1201 aaacagtaat acaaggggtg tttactagag gttgatcggg cacgtaagag gttccaactt
     1261 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     1321 agctaaggaa gctaaaatgg agaaaaaaat cacgggatat accaccgttg atatatccca
     1381 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     1441 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     1501 ttatccggcc tttattcaca ttcttgcccg cctgatgaac gctcacccgg agtttcgtat
     1561 ggccatgaaa gacggtgagc tggtgatctg ggatagtgtt cacccttgtt acaccgtttt
     1621 ccatgagcaa actgaaacgt tttcgtccct ctggagtgaa taccacgacg atttccggca
     1681 gtttctccac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     1741 taaagggttt attgagaata tgttttttgt ctcagccaat ccctgggtga gtttcaccag
     1801 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca cgatgggcaa
     1861 atattatacg caaggcgaca aggtgctgat gccgctggcg atccaggttc atcatgccgt
     1921 ttgtgatggc ttccatgtcg gccgcatgct taatgaatta caacagtact gtgatgagtg
     1981 gcagggcggg gcgtaataat actagctccg gcaaaaaaac gggcaaggtg tcaccaccct
     2041 gccctttttc tttaaaaccg aaaagattac ttcgcgtttg ccacctgacg tctaagctgt
     2101 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
     2161 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2221 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2281 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2341 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2401 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aacttgaaca
     2461 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     2521 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2581 acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2641 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2701 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2761 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2821 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2881 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtcag
     2941 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     3001 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt
     3061 tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcac caggcgttta
     3121 agggcaccaa taactgcctt aaaaaaagct agcggagtgt atactggctt actatgttgg
     3181 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     3241 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     3301 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     3361 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     3421 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     3481 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     3541 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     3601 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     3661 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     3721 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     3781 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     3841 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     3901 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     3961 agatcatctt attaatcaga taaaatattt gctcatgagc ccgaagtggc gagcccgatc
     4021 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     4081 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg
     4141 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccctcg
     4201 agccgtcaat tgtctgattc gttaccaatt attaattttt aaagtatggg caatcaattg
     4261 ctcctgttaa aattgcttta gaaatacttt ggcagcggtt tgttgtattg agtttcattt
     4321 gcgcattggt taaatggaaa gtgacagtac gctcactgca acctaatatt tttgaaatat
     4381 cccaagagct ttttccttcg catgcccacg ctaaacattc tttttctctt ttggttaaat
     4441 cgttgtttga tttattattt gctatattta tttttcgata attatcaact agagaaggaa
     4501 caattaatgg tatgttcata cacgcatgta aaaataaact atctatatag ttgtcttttt
     4561 ctgaatgtgc aaaactaagc attccgaagc cattgttagc cgtatgaata gggaaactaa
     4621 acccagtgat aagacctgat gttttcgctt ctttaattac atttggagat tttttattta
     4681 cagcattgtt ttcaaatata ttccaattaa ttggtgaatg attggagtta gaataatcta
     4741 ctataggatc atattttatt aaattagcgt catcataata ttgcctccat tttttagggt
     4801 aattatctag gattgaaata tcagatttaa ccatagaatg aggataaatg atcgcgagta
     4861 aataatattc acaatgtacc attttagtca tatcagataa gcattgatta atatcattat
     4921 tgcttctaca agctttaatt ttattaatta ttctgtatgt gtcgtcggca tttatgtttt
     4981 tcatattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     5041 ttttgcgcca ttcgatggcg cgccgcttcg tcaggccaca tagctttctt gttctgatcg
     5101 gaacgatcgt tggctgacct gtaggatcgt acaggtttac gcaagaaaat ggtttgttat
     5161 agtcgaataa atactagaga aagaggagaa ataccat
//
