LOCUS       40924_14870        7827 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDN19, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7827)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     Development of improved versatile broad-host-range vectors for use 
            in methylotrophs and other Gram-negative bacteria
  JOURNAL   Microbiology 147 (Pt 8), 2065-2075 (2001)
  PUBMED    11495985
REFERENCE   2  (bases 1 to 7827)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-DEC-2000) Microbiology, University of Washington, Box 
            357242, Seattle, WA 98195, USA
REFERENCE   3  (bases 1 to 7827)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7827)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microbiology 147 (Pt 8), 2065-2075 (2001)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-DEC-2000) Microbiology, University of Washington, Box 357242, 
            Seattle, WA 98195, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..712
                     /label=oriV
                     /note="incP origin of replication"
     primer_bind     2725..2741
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2742..2798
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(2801..2819)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2838..2854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2862..2878)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2886..2916)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2931..2952)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3280..4476
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="VKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(5755..6900)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(7175..7543)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(7576..7685)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc aggacaactg
      721 gtgagtctgg atctgaagtt cattgttgtc atggatgatc tggaccgact ggagccatcc
      781 caggtggcgg aagtgttcag gcttgtgagt gcagtagccg atctgccccg ctttacccat
      841 attctctgtt atgacaggca gattatcact catgccgttg aacatgcgct gaatatcgaa
      901 gatggcagcc gttatctcca gaaaatcatt cagcttagtt ttaaattacc ccgacctgaa
      961 gcctttgatt tacgtaatga atttcgccag cgggctgagg ctctatatca gcaaattaat
     1021 aatcaaccgc cagactctgg aatggtaagg gatctcatcg cggtgactga tacctatggt
     1081 gccgcacttt cgacgccacg ggaaatccat caggccatta attccctgat ttttctttat
     1141 ccggggatgc gggattttgt ttatttccct gatttgtgcc tgcttcagct tatacgggtg
     1201 acaaacccgg ctctgtatga ctggacagag cattacctga cagaacggtc cgtgattgaa
     1261 accggtcagg gtatgctttc tgacggagag aaagcagact tccgggaggg gcttatcaga
     1321 tgtatgaaga cgttcagggc atcaaatgca gactcgtttc tgacacttgc agactggatc
     1381 tatctcatct gcgcaaggca gaacgtgaag acggccgccc tggacctcgc ccgcgagcgc
     1441 caggcgcacg aggccggcgc gcggacccgc gccacggccc acgagcggac gccgcagcag
     1501 gagcgccaga aggccgccag agaggccgag cgcggccgtg aggcttggac gctagggcag
     1561 ggcatgaaaa agcccgtagc gggctgctac gggcgtctga cgcggtggaa agggggaggg
     1621 gatgttgtct acatggctct gctgtagtga gtgggttgcg ctccggcagc ggtcctgatc
     1681 aatcgtcacc ctttctcggt ccttcaacgt tcctgacaac gagcctcctt ttcgccaatc
     1741 catcgacaat caccgcgagt ccctgctcga acgctgcgtc cggaccggct tcgtcgaagg
     1801 cgtctatcgc ggcccgcaac agcggcgaga gcggagcctg ttcaacggtg ccgccgcgct
     1861 cgccggcatc gctgtcgccg gcctgctcct caagcacggc cccaacagtg aagtagctga
     1921 ttgtcatcag cgcattgacg gcgtccccgg ccgaaaaacc cgcctcgcag aggaagcgaa
     1981 gctgcgcgtc ggccgtttcc atctgcggtg cgcccggtcg cgtgccggca tggatgcgcg
     2041 cgccatcgcg gtaggcgagc agcgcctgcc tgaagctgcg ggcattcccg atcagaaatg
     2101 agcgccagtc gtcgtcggct ctcggcaccg aatgcgtatg attctccgcc agcatggctt
     2161 cggccagtgc gtcgagcagc gcccgcttgt tcctgaagtg ccagtaaagc gcccgctcct
     2221 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2281 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2341 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2401 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2461 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2521 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     2581 atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     2641 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     2701 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc
     2761 ggggatcctc tagagtcgac ctgcaggcat gcaagctttc cctatagtga gtcgtattag
     2821 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2881 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2941 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3001 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccgg
     3061 ctgctgaacc cccaaccgtt ccgccagttt gcgtgtcgtc agaccgtcta cgccgacctc
     3121 gttcaacagg tccagggcgg cacggatcac tgtattcggc tgcaactttg tcatgattga
     3181 cactttatca ctgataaaca taatatgtcc accaacttat cagtgataaa gaatccgcgc
     3241 gttcaatcgg accagcggag gctggtccgg aggccagacg tgaaacccaa catacccctg
     3301 atcgtaattc tgagcactgt cgcgctcgac gctgtcggca tcggcctgat tatgccggtg
     3361 ctgccgggcc tcctgcgcga tctggttcac tcgaacgacg tcaccgccca ctatggcatt
     3421 ctgctggcgc tgtatgcgtt ggtgcaattt gcctgcgcac ctgtgctggg cgcgctgtcg
     3481 gatcgtttcg ggcggcggcc aatcttgctc gtctcgctgg ccggcgccac tgtcgactac
     3541 gccatcatgg cgacagcgcc tttcctttgg gttctctata tcgggcggat cgtggccggc
     3601 atcaccgggg cgactggggc ggtagccggc gcttatattg ccgatatcac tgatggcgat
     3661 gagcgcgcgc ggcacttcgg cttcatgagc gcctgtttcg ggttcgggat ggtcgcggga
     3721 cctgtgctcg gtgggctgat gggcggtttc tccccccacg ctccgttctt cgccgcggca
     3781 gccttgaacg gcctcaattt cctgacgggc tgtttccttt tgccggagtc gcacaaaggc
     3841 gaacgccggc cgttacgccg ggaggctctc aacccgctcg cttcgttccg gtgggcccgg
     3901 ggcatgaccg tcgtcgccgc cctgatggcg gtcttcttca tcatgcaact tgtcggacag
     3961 gtgccggccg cgctttgggt cattttcggc gaggatcgct ttcactggga cgcgaccacg
     4021 atcggcattt cgcttgccgc atttggcatt ctgcattcac tcgcccaggc aatgatcacc
     4081 ggccctgtag ccgcccggct cggcgaaagg cgggcactca tgctcggaat gattgccgac
     4141 ggcacaggct acatcctgct tgccttcgcg acacggggat ggatggcgtt cccgatcatg
     4201 gtcctgcttg cttcgggtgg catcggaatg ccggcgctgc aagcaatgtt gtccaggcag
     4261 gtggatgagg aacgtcaggg gcagctgcaa ggctcactgg cggcgctcac cagcctgacc
     4321 tcgatcgtcg gacccctcct cttcacggcg atctatgcgg cttctataac aacgtggaac
     4381 gggtgggcat ggattgcagg cgctgccctc tacttgctct gcctgccggc gctgcgtcgc
     4441 gggctttgga gcggcgcagg gcaacgagcc gatcgctgat cgtggaaacg ataggcctat
     4501 gccatgcggg tcaaggcgac ttccggcaag ctatacgcgc cctaggagtg cggttggaac
     4561 gttggcccag ccagatactc ccgatcacga gcaggacgcc gatgatttga agcgcactca
     4621 gcgtctgatc caagaacaac catcctagca acacggcggt ccccgggctg agaaagccca
     4681 gtaaggaaac aactgtaggt tcgagtcgcg agatcccccg gaaccaaagg aagtaggtta
     4741 aacccgctcc gatcaggccg agccacgcca ggccgagaac attggttcct gtaggcatcg
     4801 ggattggcgg atcaaacact aaagctactg gaacgagcag aagtcctccg gccgccagtt
     4861 gccaggccgt aaaggtgagc agaggcacgg gaggttgcca cttgcgggtc agcacggttc
     4921 cgaacgccat ggaaaccgcc cccgccaggc ccgctgcgac gccgacagga tctagcgctg
     4981 cgtttggtgt caacaccaac agcgccacgc ccgcagttcc gcaaatagcc cccaggaccg
     5041 ccatcaatcg tatcgggcta cctagcagag cggcagagat gaacacgacc atcagcggct
     5101 gcacagcgcc taccgtcgcc gcgaccccgc ccggcaggcg gtagaccgaa ataaacaaca
     5161 agctccagaa tagcgaaata ttaagtgcgc cgaggatgaa gatgcgcatc caccagattc
     5221 ccgttggaat ctgtcggacg atcatcacga gcaataaacc cgccggcaac gcccgcagca
     5281 gcataccggc gacccctcgg cctcgctgtt cgggctccac gaaaacgccg gacagatgcg
     5341 ccttgtgagc gtccttgggg ccgtcctcct gtttgaagac cgacagccca atgatctcgc
     5401 cgtcgatgta ggcgccgaat gccacggcat ctcgcaaccg ttcagcgaac gcctccatgg
     5461 gctttttctc ctcgtgctcg taaacggacc cgaacatctc tggagctttc ttcagggccg
     5521 acaatcggat ctcgcggaaa tcctgcacgt cggccgctcc aagccgtcga atctgagcct
     5581 taatcacaat tgtcaatttt aatcctctgt ttatcggcag ttcgtagagc gcgccgtgcg
     5641 tcccgagcga tactgagcga agcaagtgcg tcgagcagtg cccgcttgtt cctgaaatgc
     5701 cagtaaagcg ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt
     5761 gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc
     5821 ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca
     5881 catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc
     5941 gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta
     6001 gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt
     6061 tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc
     6121 aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc
     6181 cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt
     6241 gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac
     6301 cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt
     6361 gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt
     6421 gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc
     6481 aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc
     6541 ggcctgcttg gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt
     6601 ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg
     6661 ttcgagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag
     6721 ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg
     6781 gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc
     6841 ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat
     6901 tcaccctcct tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt
     6961 tgacccgcct ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc
     7021 gtagaccgtc tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac
     7081 cagctccgcg aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg caatcacgcg
     7141 caccccccgg ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg cggaccacgc
     7201 ccatcatgac cttgccaagc tcgtcctgct tctcttcgat cttcgccagc agggcgagga
     7261 tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc
     7321 cgcccaggcg gcccaggtcg ccattgatgc gggccagctc gcggacgtgc tcatagtcca
     7381 cgacgcccgt gattttgtag ccctggccga cggccagcag gtaggccgac aggctcatgc
     7441 cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga
     7501 taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg
     7561 ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt
     7621 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     7681 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     7741 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     7801 agggttatgc agcggaaaag atccgtc
//
