LOCUS       sequence-74548-e        5200 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 5200)
  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 5200)
  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: pVRc22_4450
FEATURES             Location/Qualifiers
     source          1..5200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(685..702)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      855..941
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1033..1060
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1079..1170
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1340..1999
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1406..1425)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2141..2596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2228..2247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3282..3299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3416..3961)
                     /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(3496..3515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4033..4052
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5183..5194
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atggaactgg acgatatgaa acatgcatgg cagaccctgg aacagcgtct ggatcagcag
       61 gcagcacaga ccggtcagct gctgggtgtt gttcatgaag aaaccgttcg tagcagcctg
      121 cgtccgctgt gggttgcaca gacagcacag ctgctgtgtg cactggcact ggcaattgtt
      181 agcgcacgta gctggattcc gcataccgat cagccggttg cagttattgg tggtgttctg
      241 ctgcaggcat ggtgtatggc actggccatt agcgcaatgg ttcagctgca actgctgacc
      301 cagtttaatg ttgcaggtcc gctgctgcgt acccagcatg cactggcacg tctgcgtcgt
      361 tggcgtaccc gtgttgcacc gtggctgggt gttgcatttt gggttctgtg gattgcagtt
      421 gcagatgcac tgtggcgtgc actgacaggt cagaccctgc cgaccgattg gctggttctg
      481 aatctgctgg ttggtgttgc cggtggtatt ggcacctggc tgggttttcg tcgtctgcag
      541 cgtgcacagc atccgtggct ggaacgtatt gatcgtgcac atgcaggcac cggtgttatt
      601 cgtgcagaac gtatgctgga agaaattgca cgttttcagc gtgattaagg atccctgttt
      661 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
      721 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
      781 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
      841 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      901 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
      961 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     1021 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     1081 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1141 atgcttcaat aatattgaaa aaggaagact atgagtattc aacatttccg tgtctgctta
     1201 cataaacagt aatacaaggg gtgtttacta gaggttgatc gggcacgtaa gaggttccaa
     1261 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     1321 aggagctaag gaagctaaaa tggagaaaaa aatcacggga tataccaccg ttgatatatc
     1381 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     1441 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     1501 gttttatccg gcctttattc acattcttgc ccgcctgatg aacgctcacc cggagtttcg
     1561 tatggccatg aaagacggtg agctggtgat ctgggatagt gttcaccctt gttacaccgt
     1621 tttccatgag caaactgaaa cgttttcgtc cctctggagt gaataccacg acgatttccg
     1681 gcagtttctc cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     1741 ccctaaaggg tttattgaga atatgttttt tgtctcagcc aatccctggg tgagtttcac
     1801 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcacgatggg
     1861 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgatccagg ttcatcatgc
     1921 cgtttgtgat ggcttccatg tcggccgcat gcttaatgaa ttacaacagt actgtgatga
     1981 gtggcagggc ggggcgtaat aatactagct ccggcaaaaa aacgggcaag gtgtcaccac
     2041 cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt ttgccacctg acgtctaagc
     2101 tgtcagacca agtttactca tatatacttt agattgattt acgcgccctg tagcggcgca
     2161 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2221 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2281 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2341 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2401 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaacttga
     2461 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     2521 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2581 ttaacgttta caatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2641 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2701 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2761 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2821 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2881 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2941 caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag
     3001 caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg
     3061 ctttcgaatt tctgccattc atccgcttat tatcacttat tcaggcgtag caccaggcgt
     3121 ttaagggcac caataactgc cttaaaaaaa gctagcggag tgtatactgg cttactatgt
     3181 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     3241 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     3301 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     3361 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     3421 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     3481 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     3541 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     3601 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     3661 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3721 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     3781 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     3841 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     3901 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     3961 agaagatcat cttattaatc agataaaata tttgctcatg agcccgaagt ggcgagcccg
     4021 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     4081 gatgccggcc acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg
     4141 atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccc
     4201 tcgagccgtc aattgtctga ttcgttacca attattaatt tttaaagtat gggcaatcaa
     4261 ttgctcctgt taaaattgct ttagaaatac tttggcagcg gtttgttgta ttgagtttca
     4321 tttgcgcatt ggttaaatgg aaagtgacag tacgctcact gcaacctaat atttttgaaa
     4381 tatcccaaga gctttttcct tcgcatgccc acgctaaaca ttctttttct cttttggtta
     4441 aatcgttgtt tgatttatta tttgctatat ttatttttcg ataattatca actagagaag
     4501 gaacaattaa tggtatgttc atacacgcat gtaaaaataa actatctata tagttgtctt
     4561 tttctgaatg tgcaaaacta agcattccga agccattgtt agccgtatga atagggaaac
     4621 taaacccagt gataagacct gatgttttcg cttctttaat tacatttgga gattttttat
     4681 ttacagcatt gttttcaaat atattccaat taattggtga atgattggag ttagaataat
     4741 ctactatagg atcatatttt attaaattag cgtcatcata atattgcctc cattttttag
     4801 ggtaattatc taggattgaa atatcagatt taaccataga atgaggataa atgatcgcga
     4861 gtaaataata ttcacaatgt accattttag tcatatcaga taagcattga ttaatatcat
     4921 tattgcttct acaagcttta attttattaa ttattctgta tgtgtcgtcg gcatttatgt
     4981 ttttcatatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     5041 aggttttgcg ccattcgatg gcgcgccgct tcgtcaggcc acatagcttt cttgttctga
     5101 tcggaacgat cgttggctga cctgtaggat cgtacaggtt tacgcaagaa aatggtttgt
     5161 tatagtcgaa taaatactag agaaagagga gaaataccat
//
