LOCUS       sequence-74169-e        4552 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4552)
  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 4552)
  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..4552
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(37..54)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      207..293
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      385..412
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        431..522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             692..1351
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(758..777)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1493..1948
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1580..1599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2634..2651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2768..3313)
                     /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(2848..2867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3385..3404
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4535..4546
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 ggatccctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
       61 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
      121 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
      181 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
      241 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
      301 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
      361 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
      421 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      481 taaccctgat aaatgcttca ataatattga aaaaggaaga ctatgagtat tcaacatttc
      541 cgtgtctgct tacataaaca gtaatacaag gggtgtttac tagaggttga tcgggcacgt
      601 aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt
      661 atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcacgg gatataccac
      721 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
      781 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
      841 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaacgctca
      901 cccggagttt cgtatggcca tgaaagacgg tgagctggtg atctgggata gtgttcaccc
      961 ttgttacacc gttttccatg agcaaactga aacgttttcg tccctctgga gtgaatacca
     1021 cgacgatttc cggcagtttc tccacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     1081 cctggcctat ttccctaaag ggtttattga gaatatgttt tttgtctcag ccaatccctg
     1141 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     1201 tttcacgatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgatcca
     1261 ggttcatcat gccgtttgtg atggcttcca tgtcggccgc atgcttaatg aattacaaca
     1321 gtactgtgat gagtggcagg gcggggcgta ataatactag ctccggcaaa aaaacgggca
     1381 aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc gtttgccacc
     1441 tgacgtctaa gctgtcagac caagtttact catatatact ttagattgat ttacgcgccc
     1501 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1561 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1621 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1681 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     1741 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1801 ttccaaactt gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     1861 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1921 tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa
     1981 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2041 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2101 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2161 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2221 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2281 cctgttacca gtcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa
     2341 ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc
     2401 gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt
     2461 agcaccaggc gtttaagggc accaataact gccttaaaaa aagctagcgg agtgtatact
     2521 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
     2581 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
     2641 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     2701 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     2761 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     2821 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     2881 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     2941 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     3001 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     3061 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     3121 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     3181 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     3241 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     3301 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttgctca tgagcccgaa
     3361 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     3421 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac
     3481 agcttatcat cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc
     3541 tatgctactc cctcgagccg tcaattgtct gattcgttac caattattaa tttttaaagt
     3601 atgggcaatc aattgctcct gttaaaattg ctttagaaat actttggcag cggtttgttg
     3661 tattgagttt catttgcgca ttggttaaat ggaaagtgac agtacgctca ctgcaaccta
     3721 atatttttga aatatcccaa gagctttttc cttcgcatgc ccacgctaaa cattcttttt
     3781 ctcttttggt taaatcgttg tttgatttat tatttgctat atttattttt cgataattat
     3841 caactagaga aggaacaatt aatggtatgt tcatacacgc atgtaaaaat aaactatcta
     3901 tatagttgtc tttttctgaa tgtgcaaaac taagcattcc gaagccattg ttagccgtat
     3961 gaatagggaa actaaaccca gtgataagac ctgatgtttt cgcttcttta attacatttg
     4021 gagatttttt atttacagca ttgttttcaa atatattcca attaattggt gaatgattgg
     4081 agttagaata atctactata ggatcatatt ttattaaatt agcgtcatca taatattgcc
     4141 tccatttttt agggtaatta tctaggattg aaatatcaga tttaaccata gaatgaggat
     4201 aaatgatcgc gagtaaataa tattcacaat gtaccatttt agtcatatca gataagcatt
     4261 gattaatatc attattgctt ctacaagctt taattttatt aattattctg tatgtgtcgt
     4321 cggcatttat gtttttcata ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     4381 ccataccgcg aaaggttttg cgccattcga tggcgcgccg cttcgtcagg ccacatagct
     4441 ttcttgttct gatcggaacg atcgttggct gacctgtagg atcgtacagg tttacgcaag
     4501 aaaatggttt gttatagtcg aataaatact agagaaagag gagaaatacc at
//
