LOCUS       sequence-74539-e        4906 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 4906)
  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 4906)
  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: pVRc04_1609
FEATURES             Location/Qualifiers
     source          1..4906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(391..408)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      561..647
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      739..766
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        785..876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1046..1705
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1112..1131)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1847..2302
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1934..1953)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2988..3005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3122..3667)
                     /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(3202..3221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3739..3758
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4889..4900
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaatcgta ccaccgaaca gcgtaatgca gaagttatga aaaccattgg tctgctggat
       61 cagatgcctc gtgttgaagt tgatcacctg tttcgtgttc gtctgatgca gcgtattgaa
      121 gcaatggaag tgaaaaaaac cagctggtca gcactgcctg gtggtgcatt taatccgcgt
      181 ctggcattta tggcactgct gctgatgctg aatattgcaa gcgcactgat gctgtttatg
      241 catggtacac cgcaggcaac cggtagcagc ggtgcaattg cagaaagcct gaccgaagat
      301 tatggtggtc cggcactgag ctattatgat gatcagacca ccattgatcg ctaaggatcc
      361 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
      421 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
      481 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
      541 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
      601 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
      661 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
      721 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
      781 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      841 tgataaatgc ttcaataata ttgaaaaagg aagactatga gtattcaaca tttccgtgtc
      901 tgcttacata aacagtaata caaggggtgt ttactagagg ttgatcgggc acgtaagagg
      961 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
     1021 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc acgggatata ccaccgttga
     1081 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     1141 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1201 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaacg ctcacccgga
     1261 gtttcgtatg gccatgaaag acggtgagct ggtgatctgg gatagtgttc acccttgtta
     1321 caccgttttc catgagcaaa ctgaaacgtt ttcgtccctc tggagtgaat accacgacga
     1381 tttccggcag tttctccaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1441 ctatttccct aaagggttta ttgagaatat gttttttgtc tcagccaatc cctgggtgag
     1501 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1561 gatgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga tccaggttca
     1621 tcatgccgtt tgtgatggct tccatgtcgg ccgcatgctt aatgaattac aacagtactg
     1681 tgatgagtgg cagggcgggg cgtaataata ctagctccgg caaaaaaacg ggcaaggtgt
     1741 caccaccctg ccctttttct ttaaaaccga aaagattact tcgcgtttgc cacctgacgt
     1801 ctaagctgtc agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc
     1861 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1921 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1981 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2041 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2101 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2161 acttgaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     2221 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2281 aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2341 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2401 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2461 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2521 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2581 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2641 accagtcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     2701 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     2761 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcacc
     2821 aggcgtttaa gggcaccaat aactgcctta aaaaaagcta gcggagtgta tactggctta
     2881 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
     2941 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
     3001 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     3061 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     3121 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     3181 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     3241 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     3301 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     3361 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3421 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     3481 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     3541 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     3601 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     3661 atctcaagaa gatcatctta ttaatcagat aaaatatttg ctcatgagcc cgaagtggcg
     3721 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     3781 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     3841 tcatcgatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct
     3901 actccctcga gccgtcaatt gtctgattcg ttaccaatta ttaattttta aagtatgggc
     3961 aatcaattgc tcctgttaaa attgctttag aaatactttg gcagcggttt gttgtattga
     4021 gtttcatttg cgcattggtt aaatggaaag tgacagtacg ctcactgcaa cctaatattt
     4081 ttgaaatatc ccaagagctt tttccttcgc atgcccacgc taaacattct ttttctcttt
     4141 tggttaaatc gttgtttgat ttattatttg ctatatttat ttttcgataa ttatcaacta
     4201 gagaaggaac aattaatggt atgttcatac acgcatgtaa aaataaacta tctatatagt
     4261 tgtctttttc tgaatgtgca aaactaagca ttccgaagcc attgttagcc gtatgaatag
     4321 ggaaactaaa cccagtgata agacctgatg ttttcgcttc tttaattaca tttggagatt
     4381 ttttatttac agcattgttt tcaaatatat tccaattaat tggtgaatga ttggagttag
     4441 aataatctac tataggatca tattttatta aattagcgtc atcataatat tgcctccatt
     4501 ttttagggta attatctagg attgaaatat cagatttaac catagaatga ggataaatga
     4561 tcgcgagtaa ataatattca caatgtacca ttttagtcat atcagataag cattgattaa
     4621 tatcattatt gcttctacaa gctttaattt tattaattat tctgtatgtg tcgtcggcat
     4681 ttatgttttt catattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4741 cgcgaaaggt tttgcgccat tcgatggcgc gccgcttcgt caggccacat agctttcttg
     4801 ttctgatcgg aacgatcgtt ggctgacctg taggatcgta caggtttacg caagaaaatg
     4861 gtttgttata gtcgaataaa tactagagaa agaggagaaa taccat
//
