LOCUS       sequence-74537-e        4912 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 4912)
  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 4912)
  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: pVRc03_1198
FEATURES             Location/Qualifiers
     source          1..4912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(397..414)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      567..653
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      745..772
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        791..882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1052..1711
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1118..1137)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1853..2308
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1940..1959)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2994..3011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3128..3673)
                     /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(3208..3227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3745..3764
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4895..4906
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atggacaaag acttcgactt tgacaatatt ggtaaacgta ccccgtatcg tacaccggat
       61 aacttttttg aagaaaccca gcgcaaaatt ctggaacgta ccgttgatga acagcgtaaa
      121 aaacgtcgtc tgaaacgtat tattccgacc gttattgcag ttgcagcagt tctggcaggt
      181 attctgttta ccccgagcct gcgttatatg aataccgata ccccgagcgc aagcaatatt
      241 ctggcagttg ataaaaacaa tgttaccacc gatccggtgg acaaatggat taaagaactg
      301 agtgatgagg aactggaaga actggttagc tttagcgaaa acgatatctt cctgaattaa
      361 ggatccctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
      421 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
      481 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
      541 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
      601 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
      661 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
      721 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
      781 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      841 taaccctgat aaatgcttca ataatattga aaaaggaaga ctatgagtat tcaacatttc
      901 cgtgtctgct tacataaaca gtaatacaag gggtgtttac tagaggttga tcgggcacgt
      961 aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt
     1021 atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcacgg gatataccac
     1081 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     1141 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     1201 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaacgctca
     1261 cccggagttt cgtatggcca tgaaagacgg tgagctggtg atctgggata gtgttcaccc
     1321 ttgttacacc gttttccatg agcaaactga aacgttttcg tccctctgga gtgaatacca
     1381 cgacgatttc cggcagtttc tccacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     1441 cctggcctat ttccctaaag ggtttattga gaatatgttt tttgtctcag ccaatccctg
     1501 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     1561 tttcacgatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgatcca
     1621 ggttcatcat gccgtttgtg atggcttcca tgtcggccgc atgcttaatg aattacaaca
     1681 gtactgtgat gagtggcagg gcggggcgta ataatactag ctccggcaaa aaaacgggca
     1741 aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc gtttgccacc
     1801 tgacgtctaa gctgtcagac caagtttact catatatact ttagattgat ttacgcgccc
     1861 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1921 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1981 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2041 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     2101 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2161 ttccaaactt gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     2221 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2281 tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa
     2341 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2401 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2461 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2521 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2581 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2641 cctgttacca gtcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa
     2701 ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc
     2761 gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt
     2821 agcaccaggc gtttaagggc accaataact gccttaaaaa aagctagcgg agtgtatact
     2881 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
     2941 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
     3001 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     3061 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     3121 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     3181 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     3241 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     3301 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     3361 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     3421 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     3481 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     3541 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     3601 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     3661 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttgctca tgagcccgaa
     3721 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     3781 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac
     3841 agcttatcat cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc
     3901 tatgctactc cctcgagccg tcaattgtct gattcgttac caattattaa tttttaaagt
     3961 atgggcaatc aattgctcct gttaaaattg ctttagaaat actttggcag cggtttgttg
     4021 tattgagttt catttgcgca ttggttaaat ggaaagtgac agtacgctca ctgcaaccta
     4081 atatttttga aatatcccaa gagctttttc cttcgcatgc ccacgctaaa cattcttttt
     4141 ctcttttggt taaatcgttg tttgatttat tatttgctat atttattttt cgataattat
     4201 caactagaga aggaacaatt aatggtatgt tcatacacgc atgtaaaaat aaactatcta
     4261 tatagttgtc tttttctgaa tgtgcaaaac taagcattcc gaagccattg ttagccgtat
     4321 gaatagggaa actaaaccca gtgataagac ctgatgtttt cgcttcttta attacatttg
     4381 gagatttttt atttacagca ttgttttcaa atatattcca attaattggt gaatgattgg
     4441 agttagaata atctactata ggatcatatt ttattaaatt agcgtcatca taatattgcc
     4501 tccatttttt agggtaatta tctaggattg aaatatcaga tttaaccata gaatgaggat
     4561 aaatgatcgc gagtaaataa tattcacaat gtaccatttt agtcatatca gataagcatt
     4621 gattaatatc attattgctt ctacaagctt taattttatt aattattctg tatgtgtcgt
     4681 cggcatttat gtttttcata ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     4741 ccataccgcg aaaggttttg cgccattcga tggcgcgccg cttcgtcagg ccacatagct
     4801 ttcttgttct gatcggaacg atcgttggct gacctgtagg atcgtacagg tttacgcaag
     4861 aaaatggttt gttatagtcg aataaatact agagaaagag gagaaatacc at
//
