LOCUS       sequence-74552-e        5401 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 5401)
  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 5401)
  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: pVRc27_4265
FEATURES             Location/Qualifiers
     source          1..5401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(886..903)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1056..1142
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1234..1261
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1280..1371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1541..2200
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1607..1626)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2342..2797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2429..2448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3483..3500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3617..4162)
                     /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(3697..3716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4234..4253
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5384..5395
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgacaggtc atccggatgt tgcagaaatt gcagatctgg cagaaggtct gctgccgacc
       61 acccgtacca ccgaagttcg tcagcatctg gaaagctgtg aactgtgtgc agatgtttat
      121 gcaagcctga ccgaaattca gggtctgctg ggcaccctgc ctgcaccggc accgatgccg
      181 gatgatgttg cagcacgtat tgatgcagca ctggcagcag aaccgcctct gggtattgca
      241 gatggcaccc gtgttagccg tgaaaccagc acaccggcag atcgtccggc aggtcatgca
      301 cgtccgagca gcacaggtcc gggtcgtaaa gatcgtcgtc gtggtggtcg tcgccgtatt
      361 gcagttctgg gtgcagttgc agcagcagca gccattggta ttggtagcgt tgttgttagc
      421 agtctgaccg aagatagcag cagcggtaat accgcacgtg aacagcagac cgcaattgca
      481 gataccttta gcgaaggtcg tctgaaagat cgtgttacca atctggttgc agatggtagc
      541 gcagaaaatg gtagccgtac accgcgtagc tttggtatgg aaagcgaaaa tggtggtgaa
      601 accgcagaaa accatgtttt taaacagccg accgttccgg aatgtattcg taaaggtatt
      661 ggtcgtgatg atgcagttat tgcaaccgaa ccgggtgttt ataaaggtcg tgaagcactg
      721 ctggttgttc tgccggatgc aaccaatgat acccaggtta ccgcatatat tgttgaaacc
      781 gcatgtgttg atcagcctgc agttggtaaa gcaaaaattc tgctggaaca tagctatgca
      841 cgtagctaag gatccctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
      901 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
      961 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1021 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1081 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1141 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1201 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1261 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1321 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagac tatgagtatt
     1381 caacatttcc gtgtctgctt acataaacag taatacaagg ggtgtttact agaggttgat
     1441 cgggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt
     1501 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcacggg
     1561 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     1621 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     1681 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     1741 gaacgctcac ccggagtttc gtatggccat gaaagacggt gagctggtga tctgggatag
     1801 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcgt ccctctggag
     1861 tgaataccac gacgatttcc ggcagtttct ccacatatat tcgcaagatg tggcgtgtta
     1921 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt ttgtctcagc
     1981 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     2041 cgcccccgtt ttcacgatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     2101 ggcgatccag gttcatcatg ccgtttgtga tggcttccat gtcggccgca tgcttaatga
     2161 attacaacag tactgtgatg agtggcaggg cggggcgtaa taatactagc tccggcaaaa
     2221 aaacgggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg
     2281 tttgccacct gacgtctaag ctgtcagacc aagtttactc atatatactt tagattgatt
     2341 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2461 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2521 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     2581 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2641 ggactcttgt tccaaacttg aacaacactc aaccctatct cgggctattc ttttgattta
     2701 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2761 aacgcgaatt ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct
     2821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3121 tctgctaatc ctgttaccag tcaggcattt gagaagcaca cggtcacact gcttccggta
     3181 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
     3241 ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta
     3301 ttcaggcgta gcaccaggcg tttaagggca ccaataactg ccttaaaaaa agctagcgga
     3361 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     3421 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     3481 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     3541 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     3601 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     3661 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3721 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     3781 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     3841 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     3901 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     3961 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     4021 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     4081 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     4141 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttgctcat
     4201 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     4261 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc
     4321 atgtttgaca gcttatcatc gatgcataat gtgcctgtca aatggacgaa gcagggattc
     4381 tgcaaaccct atgctactcc ctcgagccgt caattgtctg attcgttacc aattattaat
     4441 ttttaaagta tgggcaatca attgctcctg ttaaaattgc tttagaaata ctttggcagc
     4501 ggtttgttgt attgagtttc atttgcgcat tggttaaatg gaaagtgaca gtacgctcac
     4561 tgcaacctaa tatttttgaa atatcccaag agctttttcc ttcgcatgcc cacgctaaac
     4621 attctttttc tcttttggtt aaatcgttgt ttgatttatt atttgctata tttatttttc
     4681 gataattatc aactagagaa ggaacaatta atggtatgtt catacacgca tgtaaaaata
     4741 aactatctat atagttgtct ttttctgaat gtgcaaaact aagcattccg aagccattgt
     4801 tagccgtatg aatagggaaa ctaaacccag tgataagacc tgatgttttc gcttctttaa
     4861 ttacatttgg agatttttta tttacagcat tgttttcaaa tatattccaa ttaattggtg
     4921 aatgattgga gttagaataa tctactatag gatcatattt tattaaatta gcgtcatcat
     4981 aatattgcct ccatttttta gggtaattat ctaggattga aatatcagat ttaaccatag
     5041 aatgaggata aatgatcgcg agtaaataat attcacaatg taccatttta gtcatatcag
     5101 ataagcattg attaatatca ttattgcttc tacaagcttt aattttatta attattctgt
     5161 atgtgtcgtc ggcatttatg tttttcatat tcaccaccct gaattgactc tcttccgggc
     5221 gctatcatgc cataccgcga aaggttttgc gccattcgat ggcgcgccgc ttcgtcaggc
     5281 cacatagctt tcttgttctg atcggaacga tcgttggctg acctgtagga tcgtacaggt
     5341 ttacgcaaga aaatggtttg ttatagtcga ataaatacta gagaaagagg agaaatacca
     5401 t
//
