LOCUS       40924_46548        6372 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWH1274, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6372)
  AUTHORS   Wang Y, Chen Y, Zhou Q, Huang S, Ning K, Xu J, Kalin RM, Rolfe S, 
            Huang WE.
  TITLE     A culture-independent approach to unravel uncultured bacteria and 
            functional genes in a complex microbial community
  JOURNAL   PLoS ONE 7 (10), E47530 (2012)
  PUBMED    23082176
REFERENCE   2  (bases 1 to 6372)
  AUTHORS   Wang Y, Huang W.
  TITLE     Development of a toolbox for the dissection of naphthalene 
            degradation genes from unculturable microorganisms in complex 
            environment
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6372)
  AUTHORS   Wang Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUL-2011) Kroto Research Institute, University of 
            Sheffield, Broad Lane, Sheffield, South Yorkshire S3 7HQ, United 
            Kingdom
REFERENCE   4  (bases 1 to 6372)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6372)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2012"; volume: "7"; issue: "10"; pages: "E47530"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (20-JUL-2011) Kroto Research Institute, University of Sheffield, 
            Broad Lane, Sheffield, South Yorkshire S3 7HQ, United Kingdom"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..38
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             86..1273
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    4219..4359
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4545..5133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5307..6164)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6165..6269)
                     /label=AmpR promoter
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
      241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
      301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
      361 cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
      421 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
      481 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
      541 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
      601 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat
      661 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
      721 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
      781 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
      841 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
      901 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
      961 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     1021 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     1081 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc
     1141 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
     1201 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     1261 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
     1321 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
     1381 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg
     1441 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     1501 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     1561 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     1621 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     1681 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     1741 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     1801 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     1861 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     1921 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     1981 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     2041 gaccacgctg atgagcttta ccgcagagac catgacctaa agcaagaatt ttcaagcctt
     2101 gctaagactg ttgatcggtc gggggcgcag ctaatccgtg acttcatgcg tgactttgtt
     2161 aagaagcaac aggaagccgc agactatgac aaatggttca agcagcaggt acagatcggc
     2221 ttaaatgaag ccaatgcagg caagttgatt ccacatgagg acgtaaaggc agagtttgcc
     2281 gctagacgtg ctgcaacatt ggcgaaactg gcagcgaaga atgaagattg agtggactga
     2341 aacagctcgc caagatcgta gaaatatcta tgattatctt gaagaacgca accctatagc
     2401 agctattgaa attgatgatt taattgaaga aaagacagat ttacttgttg ataatcgact
     2461 gatggggcgc acaggcagac agaaagatac tagggagtta gtgatacatc cgcattatgt
     2521 ggttgtatat gacatcactg atataatacg gatactcaga gtgctacaca catcgcagga
     2581 gtggtcatga cttactcatg tactttggat tatttagtgt tataaaatcc tgatttataa
     2641 attttttttg ttaaaaaaga taaaagcccc ttgcaattgc ttggggcttt accgtaattt
     2701 atggggtaca gatcttcgat actgacatat cggcaatcga aagcattaan gtttgacgac
     2761 cgctaatgat ttcaccacan gggcttaatg tacctgtctt aaattctaag gttttaactc
     2821 gctttgtcaa gcatagaccc caaaaattta gccaatgtct gtaactcaat ctgtccatgt
     2881 gtgggtgatg aggtacagtg acgctagcac acatcggaaa aacgctatta ctaggggaac
     2941 tgaacagagt agcggacgca atgagtagtc atttaattgg cggttatgag cgtgttcagg
     3001 cggtgctatc aatcgtaatc ataacagtgg cagcttgata cagtgatgtc atccctgatg
     3061 cgaaagcgac cgaccgacgg tacatcgaat gggaatactt tagggtgatt tttaagaatc
     3121 gctctagggt gagtatttcc cattcagctc tgctccctcc ctctggtact ttaatcaaaa
     3181 gcactactaa acatatgttt ttaaataaaa aatattgata tagagataat attagtaaga
     3241 ataattaaac aattgaatat agataaatca ttgttaaata aagattaatt attaaaatga
     3301 atgtatactt atatataaat caatgattta aaatatttga taaagaaaac ttttcaaaaa
     3361 aaatataatt gagattgtgt catttcggtc aattcttaat atgttccacg caagttttag
     3421 ctatggtgct aaacagaaat ttgctgaaaa agaacttttc actgaactgg ttaaaatgta
     3481 agcagcntga gagccgccaa aaattttaaa aacaaaccgc cttaatcatc ttcaaaaaat
     3541 acctctaaaa cctcaccatt tgcgttttaa gacccatatt tcatcctgcc cttatgttcc
     3601 catgctgata gctataaagt gtctgtaatc gcttcctatg acgttntagg ctgttgataa
     3661 cttttggaac aacgcaaaat gttaaaatcc gatcattttt taacctagtt attttcgtta
     3721 cagggtaact ttggtagtat tatttcaata tttagttgtt acaggataac aaaatatatg
     3781 ttacagggta attaatttag cgttacagga taactatagt tacagggtaa cgttaaatag
     3841 ttatcctgta atttaaaaat agttgcagga tagcataaat ttttacaaat aaattttata
     3901 aaaagttatc ctgtaactaa aaacaatgtt atcctgtaac tatgatatac tatatgggag
     3961 ttaaagacat gaaagacccg aacgatcaga aaacacagga catgctaaaa gagccgccaa
     4021 aaaccaatgc cgaacgacaa aaagcatatc gtgaaaaacg caaaagcctt gatagtcctg
     4081 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4141 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4201 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     4261 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     4321 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     4381 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4741 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4981 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5101 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5221 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5281 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5341 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5401 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5461 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5521 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5581 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     5641 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5701 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5761 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5821 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5881 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     5941 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6001 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6061 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6121 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6181 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6241 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6301 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6361 cgtcttcaag aa
//
