LOCUS       sequence-74549-e        5158 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5158)
  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 5158)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pVRc25_1643_cl43
FEATURES             Location/Qualifiers
     source          1..5158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(643..660)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      813..899
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      991..1018
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1037..1128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1298..1957
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1364..1383)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2099..2554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2186..2205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3240..3257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3374..3919)
                     /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(3454..3473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3991..4010
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5141..5152
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaccaccg atagcaactt taatgatccg tgtcgtcgtc agtttagccg tgatctgccg
       61 cagaaaatgg cacgtcatac caatgaagca accggtgcaa tggatatggt taaacgtgat
      121 cgttctgaac tgctgagcgc atatctggat ggtgaagtta ccgcagcaga acgtcgtcag
      181 gttgaagatt ggctggcaaa tgatgttgca gttcagcgtc tgtatagccg tctgctgaaa
      241 ctgcgtcagg gtattcgtac catgccgatt ccgaccgcac agcagagtcc ggaaaccacc
      301 gcagaacagg tttttgcaaa agttaatcgt cgtagccgtc tggcatggaa actgggtggt
      361 gcagcagttg cagcatgtgt tattggtgca gttaccaatt ggctgcctgg tcgtcagacc
      421 ggtattccgc agctggcaca gcagccgcaa gaacagccga cccaggcagt taccacaccg
      481 gatgcactgc cgatgattgc actgaataat ccggttattg aaattccgaa agcagcagtt
      541 gccagcccga ccaaatttat ccagccgcag ccgcagctgg gtgaaattcc gcctgatatt
      601 aactaaggat ccctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc
      661 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
      721 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
      781 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
      841 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
      901 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
      961 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
     1021 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1081 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagactat gagtattcaa
     1141 catttccgtg tctgcttaca taaacagtaa tacaaggggt gtttactaga ggttgatcgg
     1201 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     1261 tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcacgggata
     1321 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     1381 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     1441 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     1501 cgctcacccg gagtttcgta tggccatgaa agacggtgag ctggtgatct gggatagtgt
     1561 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcgtccc tctggagtga
     1621 ataccacgac gatttccggc agtttctcca catatattcg caagatgtgg cgtgttacgg
     1681 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgttttttg tctcagccaa
     1741 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     1801 ccccgttttc acgatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     1861 gatccaggtt catcatgccg tttgtgatgg cttccatgtc ggccgcatgc ttaatgaatt
     1921 acaacagtac tgtgatgagt ggcagggcgg ggcgtaataa tactagctcc ggcaaaaaaa
     1981 cgggcaaggt gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgttt
     2041 gccacctgac gtctaagctg tcagaccaag tttactcata tatactttag attgatttac
     2101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2221 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2281 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     2341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2401 ctcttgttcc aaacttgaac aacactcaac cctatctcgg gctattcttt tgatttataa
     2461 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2521 gcgaatttta acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt
     2581 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2881 gctaatcctg ttaccagtca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     2941 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     3001 acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc
     3061 aggcgtagca ccaggcgttt aagggcacca ataactgcct taaaaaaagc tagcggagtg
     3121 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     3181 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     3241 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     3301 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     3361 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     3421 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
     3481 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     3541 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     3601 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     3661 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     3721 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     3781 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     3841 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     3901 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tgctcatgag
     3961 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     4021 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg
     4081 tttgacagct tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     4141 aaaccctatg ctactccctc gagccgtcaa ttgtctgatt cgttaccaat tattaatttt
     4201 taaagtatgg gcaatcaatt gctcctgtta aaattgcttt agaaatactt tggcagcggt
     4261 ttgttgtatt gagtttcatt tgcgcattgg ttaaatggaa agtgacagta cgctcactgc
     4321 aacctaatat ttttgaaata tcccaagagc tttttccttc gcatgcccac gctaaacatt
     4381 ctttttctct tttggttaaa tcgttgtttg atttattatt tgctatattt atttttcgat
     4441 aattatcaac tagagaagga acaattaatg gtatgttcat acacgcatgt aaaaataaac
     4501 tatctatata gttgtctttt tctgaatgtg caaaactaag cattccgaag ccattgttag
     4561 ccgtatgaat agggaaacta aacccagtga taagacctga tgttttcgct tctttaatta
     4621 catttggaga ttttttattt acagcattgt tttcaaatat attccaatta attggtgaat
     4681 gattggagtt agaataatct actataggat catattttat taaattagcg tcatcataat
     4741 attgcctcca ttttttaggg taattatcta ggattgaaat atcagattta accatagaat
     4801 gaggataaat gatcgcgagt aaataatatt cacaatgtac cattttagtc atatcagata
     4861 agcattgatt aatatcatta ttgcttctac aagctttaat tttattaatt attctgtatg
     4921 tgtcgtcggc atttatgttt ttcatattca ccaccctgaa ttgactctct tccgggcgct
     4981 atcatgccat accgcgaaag gttttgcgcc attcgatggc gcgccgcttc gtcaggccac
     5041 atagctttct tgttctgatc ggaacgatcg ttggctgacc tgtaggatcg tacaggttta
     5101 cgcaagaaaa tggtttgtta tagtcgaata aatactagag aaagaggaga aataccat
//
