LOCUS       sequence-74540-e        5212 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 5212)
  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 5212)
  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: pVRc11_987
FEATURES             Location/Qualifiers
     source          1..5212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(697..714)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      867..953
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1045..1072
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1091..1182
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1352..2011
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1418..1437)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2153..2608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2240..2259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3294..3311)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3428..3973)
                     /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(3508..3527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4045..4064
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5195..5206
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaacaaac acccggataa taatctgctg gaagcctatg caagcggtag cattgatgca
       61 gttagcggtc tggttgttgc aacccatctg gaaacctgta gcaaatgtcg tgcctatgtt
      121 aatcaggttg aagcaagcca ggcaaatacc gttagcgaaa gcccgagcga atatagtccg
      181 gaatttgatg atatgctgaa tgacattatt aacgccgaac cggtgaatga taatgtggtt
      241 attcaggata ccgcctttgt taatgttgca ggtaaatcat ttgaactgcc gaaaaccctg
      301 gttcgtttta gcgatctggt tggtagctgg cgtagctatg gtggtaaagt ttttagcgca
      361 cagattgatc tgggtgaaga tgcacgtgtt agcctgatgt atattggtga aaatgttcag
      421 attccgcagc atacccatcg tggtctggaa agcaccctgg ttctgcatgg tggttttagt
      481 gatgaagatg gtcagtatga agagggtgat ctgatggttc gtgatgcaag cgttaaacat
      541 tcaccgttta cccaagaggg tgaagattgt ctgtgtctga ccgttctgac cgaaccgatg
      601 atttttaccc agggtgttgc acgcattttt aacctgtttg gtaaaggtct gtatccgtaa
      661 ggatccctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
      721 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
      781 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
      841 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
      901 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
      961 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     1021 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     1081 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1141 taaccctgat aaatgcttca ataatattga aaaaggaaga ctatgagtat tcaacatttc
     1201 cgtgtctgct tacataaaca gtaatacaag gggtgtttac tagaggttga tcgggcacgt
     1261 aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt
     1321 atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcacgg gatataccac
     1381 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     1441 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     1501 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaacgctca
     1561 cccggagttt cgtatggcca tgaaagacgg tgagctggtg atctgggata gtgttcaccc
     1621 ttgttacacc gttttccatg agcaaactga aacgttttcg tccctctgga gtgaatacca
     1681 cgacgatttc cggcagtttc tccacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     1741 cctggcctat ttccctaaag ggtttattga gaatatgttt tttgtctcag ccaatccctg
     1801 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     1861 tttcacgatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgatcca
     1921 ggttcatcat gccgtttgtg atggcttcca tgtcggccgc atgcttaatg aattacaaca
     1981 gtactgtgat gagtggcagg gcggggcgta ataatactag ctccggcaaa aaaacgggca
     2041 aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc gtttgccacc
     2101 tgacgtctaa gctgtcagac caagtttact catatatact ttagattgat ttacgcgccc
     2161 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2221 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2281 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2341 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     2401 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2461 ttccaaactt gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     2521 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2581 tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa
     2641 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2701 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2761 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2821 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2881 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2941 cctgttacca gtcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa
     3001 ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc
     3061 gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt
     3121 agcaccaggc gtttaagggc accaataact gccttaaaaa aagctagcgg agtgtatact
     3181 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
     3241 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
     3301 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     3361 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     3421 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     3481 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     3541 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     3601 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     3661 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     3721 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     3781 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     3841 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     3901 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     3961 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttgctca tgagcccgaa
     4021 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     4081 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac
     4141 agcttatcat cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc
     4201 tatgctactc cctcgagccg tcaattgtct gattcgttac caattattaa tttttaaagt
     4261 atgggcaatc aattgctcct gttaaaattg ctttagaaat actttggcag cggtttgttg
     4321 tattgagttt catttgcgca ttggttaaat ggaaagtgac agtacgctca ctgcaaccta
     4381 atatttttga aatatcccaa gagctttttc cttcgcatgc ccacgctaaa cattcttttt
     4441 ctcttttggt taaatcgttg tttgatttat tatttgctat atttattttt cgataattat
     4501 caactagaga aggaacaatt aatggtatgt tcatacacgc atgtaaaaat aaactatcta
     4561 tatagttgtc tttttctgaa tgtgcaaaac taagcattcc gaagccattg ttagccgtat
     4621 gaatagggaa actaaaccca gtgataagac ctgatgtttt cgcttcttta attacatttg
     4681 gagatttttt atttacagca ttgttttcaa atatattcca attaattggt gaatgattgg
     4741 agttagaata atctactata ggatcatatt ttattaaatt agcgtcatca taatattgcc
     4801 tccatttttt agggtaatta tctaggattg aaatatcaga tttaaccata gaatgaggat
     4861 aaatgatcgc gagtaaataa tattcacaat gtaccatttt agtcatatca gataagcatt
     4921 gattaatatc attattgctt ctacaagctt taattttatt aattattctg tatgtgtcgt
     4981 cggcatttat gtttttcata ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     5041 ccataccgcg aaaggttttg cgccattcga tggcgcgccg cttcgtcagg ccacatagct
     5101 ttcttgttct gatcggaacg atcgttggct gacctgtagg atcgtacagg tttacgcaag
     5161 aaaatggttt gttatagtcg aataaatact agagaaagag gagaaatacc at
//
