LOCUS       sequence-74555-e        5248 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 5248)
  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 5248)
  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: pVRc33_375
FEATURES             Location/Qualifiers
     source          1..5248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(733..750)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      903..989
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1081..1108
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1127..1218
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1388..2047
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1454..1473)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2189..2644
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2276..2295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3330..3347)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3464..4009)
                     /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(3544..3563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4081..4100
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5231..5242
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgatggccc tgagcaaaaa aatgctggaa caagaaccga gcgaaattga actgctgctg
       61 ccgtggcatg ctgcaggcac cctgaatgca cgtgatgcac gtcgtgttga agatgcactg
      121 gcacgtgatc cggaactggc aaaacagtat gcagcaattc gtggtgaata tgaagaaacc
      181 attcatctga atgaaagcct gggtgcaccg agcgcacgtg caatgcagaa actgtttggt
      241 gcaattgatg cagaaccggc acgtgaaacc ggtagcctgc cgctgagcgc acgtattgca
      301 accttttttg caagcctgag tccgcgtacc ctggcatggt cagccagcct gggtgccgtt
      361 gcactggttc tgcaggcagg tattattggt gcagttctga tgaaaaccca gccgaccacc
      421 tttcagaccg caagcctgag caccagcgca ccgattaccc gtgaactggg tgcagcagtt
      481 gcaccggcac gtgcactggt tcgttttaca ccggaagcac gtgttgcaga tattaccgca
      541 ctgctggata gctatcaggc aagcattatt ggtgatgcaa aaggtggtat gtttcgcctg
      601 cagtttgata aagcaatgag ccaggatgaa ctggcaagcc tgctgggtcg tatgcagcgt
      661 gaaaaatttg ttaatctggc agttgcagca ccgtaaggat ccctgttttg gcggatgaga
      721 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
      781 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
      841 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
      901 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      961 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     1021 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     1081 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     1141 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1201 tattgaaaaa ggaagactat gagtattcaa catttccgtg tctgcttaca taaacagtaa
     1261 tacaaggggt gtttactaga ggttgatcgg gcacgtaaga ggttccaact ttcaccataa
     1321 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     1381 agctaaaatg gagaaaaaaa tcacgggata taccaccgtt gatatatccc aatggcatcg
     1441 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1501 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1561 ctttattcac attcttgccc gcctgatgaa cgctcacccg gagtttcgta tggccatgaa
     1621 agacggtgag ctggtgatct gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1681 aactgaaacg ttttcgtccc tctggagtga ataccacgac gatttccggc agtttctcca
     1741 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1801 tattgagaat atgttttttg tctcagccaa tccctgggtg agtttcacca gttttgattt
     1861 aaacgtggcc aatatggaca acttcttcgc ccccgttttc acgatgggca aatattatac
     1921 gcaaggcgac aaggtgctga tgccgctggc gatccaggtt catcatgccg tttgtgatgg
     1981 cttccatgtc ggccgcatgc ttaatgaatt acaacagtac tgtgatgagt ggcagggcgg
     2041 ggcgtaataa tactagctcc ggcaaaaaaa cgggcaaggt gtcaccaccc tgcccttttt
     2101 ctttaaaacc gaaaagatta cttcgcgttt gccacctgac gtctaagctg tcagaccaag
     2161 tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg
     2221 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2281 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2341 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2401 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2461 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaacttgaac aacactcaac
     2521 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2581 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     2641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2881 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtca ggcatttgag
     3001 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     3061 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc
     3121 tgccattcat ccgcttatta tcacttattc aggcgtagca ccaggcgttt aagggcacca
     3181 ataactgcct taaaaaaagc tagcggagtg tatactggct tactatgttg gcactgatga
     3241 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
     3301 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
     3361 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
     3421 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
     3481 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
     3541 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
     3601 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
     3661 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
     3721 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
     3781 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
     3841 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt
     3901 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
     3961 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     4021 tattaatcag ataaaatatt tgctcatgag cccgaagtgg cgagcccgat cttccccatc
     4081 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     4141 gatgcgtccg gcgtagagga tctgctcatg tttgacagct tatcatcgat gcataatgtg
     4201 cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccctc gagccgtcaa
     4261 ttgtctgatt cgttaccaat tattaatttt taaagtatgg gcaatcaatt gctcctgtta
     4321 aaattgcttt agaaatactt tggcagcggt ttgttgtatt gagtttcatt tgcgcattgg
     4381 ttaaatggaa agtgacagta cgctcactgc aacctaatat ttttgaaata tcccaagagc
     4441 tttttccttc gcatgcccac gctaaacatt ctttttctct tttggttaaa tcgttgtttg
     4501 atttattatt tgctatattt atttttcgat aattatcaac tagagaagga acaattaatg
     4561 gtatgttcat acacgcatgt aaaaataaac tatctatata gttgtctttt tctgaatgtg
     4621 caaaactaag cattccgaag ccattgttag ccgtatgaat agggaaacta aacccagtga
     4681 taagacctga tgttttcgct tctttaatta catttggaga ttttttattt acagcattgt
     4741 tttcaaatat attccaatta attggtgaat gattggagtt agaataatct actataggat
     4801 catattttat taaattagcg tcatcataat attgcctcca ttttttaggg taattatcta
     4861 ggattgaaat atcagattta accatagaat gaggataaat gatcgcgagt aaataatatt
     4921 cacaatgtac cattttagtc atatcagata agcattgatt aatatcatta ttgcttctac
     4981 aagctttaat tttattaatt attctgtatg tgtcgtcggc atttatgttt ttcatattca
     5041 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     5101 attcgatggc gcgccgcttc gtcaggccac atagctttct tgttctgatc ggaacgatcg
     5161 ttggctgacc tgtaggatcg tacaggttta cgcaagaaaa tggtttgtta tagtcgaata
     5221 aatactagag aaagaggaga aataccat
//
