LOCUS       V002746                 6279 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002746
VERSION     V002746
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6279)
  AUTHORS   Nallapareddy SR, Singh KV, Murray BE.
  TITLE     Construction of Improved Temperature-Sensitive and Mobilizable
            Vectors and Their Use for Constructing Mutations in the
            Adhesin-Encoding acm Gene of Poorly Transformable Clinical
            Enterococcus faecium Strains
  JOURNAL   Appl. Environ. Microbiol. 72 (1), 334-345 (2006)
   PUBMED   16391062
REFERENCE   2  (bases 1 to 6279)
  AUTHORS   Nallapareddy SR, Singh KV, Murray BE.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-SEP-2005) Infectious Diseases, Univ. of Texas Med.
            School at Houston, 6431 Fannin St, MSB 2.112, Houston, TX 77030, USA
REFERENCE   3  (bases 1 to 6279)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6279)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2006"; volume: "72"; issue: "1"; pages:
            "334-345"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (18-SEP-2005) Infectious Diseases, Univ. of Texas Med. School at
            Houston, 6431 Fannin St, MSB 2.112, Houston, TX 77030, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..61
                     /note="multiple cloning site I; MCS-I"
     CDS             195..842
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     misc_feature    1007..1076
                     /note="multiple cloning site II; MCS-II"
     CDS             complement(1479..1490)
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             1624..1833
                     /codon_start=1
                     /gene="orfB"
                     /product="OrfB"
                     /label="orfB"
                     /note="portion of pWV01 based temperature-sensitive
                     gram-positive replicon derived from pTV1-OK"
                     /protein_id="ABB20801.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     gene            1624..1833
                     /gene="orfB"
                     /label="orfB"
     CDS             1874..2035
                     /codon_start=1
                     /product="OrfC"
                     /label="OrfC"
                     /note="portion of pWV01 based temperature-sensitive
                     gram-positive replicon derived from pTV1-OK"
                     /protein_id="ABB20802.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     CDS             2102..2800
                     /codon_start=1
                     /gene="repA(ts)"
                     /product="RepA(ts)"
                     /label="repA(ts)"
                     /note="portion of pWV01 based temperature-sensitive
                     gram-positive replicon derived from pTV1-OK"
                     /protein_id="ABB20803.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDLYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            2102..2800
                     /gene="repA(ts)"
                     /label="repA(ts)"
     CDS             2797..2946
                     /codon_start=1
                     /gene="orfD"
                     /product="OrfD"
                     /label="orfD"
                     /note="portion of pWV01 based temperature-sensitive
                     gram-positive replicon derived from pTV1-OK"
                     /protein_id="ABB20804.1"
                     /translation="MTNKEKELFAENEELKKEIKDLKERIERYREMEVELSTTIDLLRG
                     GIIE"
     gene            2797..2946
                     /gene="orfD"
                     /label="orfD"
     CDS             3173..4078
                     /codon_start=1
                     /gene="aph(2'')-Id"
                     /product="APH(2'')-Id"
                     /label="aph(2'')-Id"
                     /note="aminoglycoside modifying enzyme; derived from
                     Enterococcus casseliflavus in GenBank Accession Number
                     AF016483"
                     /protein_id="ABB20805.1"
                     /translation="MRTYTFDQVEKAIEQLYPDFTINTIEISGEGNDCIAYEINRDFIF
                     KFPKHSRGSTNLFNEVNILKRIHNKLPLPIPEVVFTGMPSETYQMSFAGFTKIKGVPLT
                     PLLLNNLPKQSQNQAAKDLARFLSELHSINISGFKSNLVLDFREKINEDNKKIKKLLSR
                     ELKGPQMKIVDDFYRDILENEIYFKYYPCLIHNDFSSDHILFDTEKNTICGIIDFGDAA
                     ISDPDNDFISLMEDDEEYGMEFVSKILNHYKHKDIPTVLEKYRMKEKYWSFEKIIYGKE
                     YGYMDWYEEGLNEIRSIKIK"
     gene            3173..4078
                     /gene="aph(2'')-Id"
                     /label="aph(2'')-Id"
     oriT            4172..4838
                     /note="oriT of Tn916 derived from pHS1; facilitates
                     conjugative transfer from the transformable E. faecalis
                     strain JH2Sm::Tn916 to E. faecium"
     rep_origin      complement(5036..5624)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 tgtacaattg ctagcgtacg gccgtcgacg tccggatcct cgagctctag atcttaagct
       61 tgattttcgt tcgtgaatac atgttataat aactataact aataacgtaa cgtaaaattt
      121 gtttgatttg tccaaaatat ggtataatag gtactaatca aaatagtgag gtaaccaatt
      181 cggaggcata tcaaatgaac tttaataaaa ttgatttaga caattggaag agaaaagaga
      241 tatttaatca ttatttgaac caacaaacga cttttagtat aaccacagaa attgatatta
      301 gtgttttata ccgaaacata aaacaagaag gatataaatt ttaccctgca tttattttct
      361 tagtgacaag ggtgataaac tcaaatacag cttttagaac tggttacaat agcgacggag
      421 agttaggtta ttgggataag ttagagccac tttatacaat ttttgatggt gtatctaaaa
      481 cattctctgg tatttggact cctgtaaaga atgacttcaa agagttttat gatttatacc
      541 tttctgatgt agagaaatat aatggttcgg ggaaattgtt tcccaaaaca cctatacctg
      601 aaaatgcttt ttctctttct attattccat ggacttcatt tactgggttt aacttaaata
      661 tcaataataa tagtaattac cttctaccca ttattacagc aggaaaattc attaataaag
      721 gtaattcaat atatttaccg ctatctttac aggtacatca ttctgtttgt gatggttatc
      781 atgcaggatt gtttatgaac tctattcagg aattgtcaga taggcctaat gactggcttt
      841 tataatatac caggtaatgg actaatgaaa atgtaaattt aactataaac tatttaaata
      901 acagattaaa aaaattataa aaaaattgaa aaaatggtgg aaacactttc ttcaattttt
      961 ttgttttatt atttaatatt tgggaaatat tcattctaat tggtaagcgc gccatgggcc
     1021 ctgcagcatg catatgcatc cggaattccc gggatcgatc gaattcggta cccggggatc
     1081 gttaaattta tactgcaatc ggatgcgatt attgaataaa agatatgaga gatttatcta
     1141 atttcttttt tcttgtaaaa aaagaaagtt cttaaaggtt ttatagtttt ggtcgtagag
     1201 cacacggttt aacgacttaa ttacgaagta aataagtcta gtgtgttaga ctttatgaaa
     1261 tctatatacg tttatatata tttattatcc gattttttat taaaacgtct caaaatcgtt
     1321 tctgagacgt tttagcgttt atttcgttta gttatcggca taatcgttaa aacaggcgtt
     1381 atcgtagcgt aaaagccctt gagcgtagcg tggctttgca gcgaagatgt tgtctgttag
     1441 attatgaaag ccgatgactg aatgaaataa taagcgcagc gcccttctat ttcggttgga
     1501 ggaggctcaa gggagtatga gggaatgaaa ttccctcatg ggtttgattt taaaaattgc
     1561 ttgcaatttt gccgagcggt agcgctggaa aatttttgaa aaaaatttgg aatttggaaa
     1621 aaaatggggg gaaaggaagc gaattttgct tccgtactac gaccccccat taagtgccga
     1681 gtgccaattt ttgtgccaaa aacgctctat cccaactggc tcaagggttt aaggggtttt
     1741 tcaatcgcca acgaatcgcc aacgttttcg ccaacgtttt ttataaatct atatttaagt
     1801 agctttattg ttgtttttat gattacaaag tgatacacta actttataaa attatttgat
     1861 tggagttttt taaatggtga tttcagaatc gaaaaaaaga gttatgattt ctctgacaaa
     1921 agagcaagat aaaaaattaa cagatatggc gaaacaaaaa ggtttttcaa aatctgcggt
     1981 tgcggcgtta gctatagaag aatatgcaag aaaggaatca gaacaaaaaa aataagcgaa
     2041 agctcgcgtt tttagaagga tacgagtttt cgctacttgt ttttgataag gtaattatat
     2101 catggctatt aaaaatacta aagctagaaa ttttggattt ttattatatc ctgactcaat
     2161 tcctaatgat tggaaagaaa aattagagag tttgggcgta tctatggctg tcagtccttt
     2221 acacgatatg gacgaaaaaa aagataaaga tacatggaat aatagtaata ttatacaaaa
     2281 tggaaagcac tataaaaaac cacactatca cgttatatat attgcacgaa atcctgtaac
     2341 aatagaaagc gttaggaaca agattaagcg aaaattgggg aatagttcag ttgctcatgt
     2401 tgagatactt gattatatca aaggttcata tgaatatttg actcatgaat caaaggacgc
     2461 tattgctaag aataaacata tatacgacaa aaaagatatt ttgaacatta atgattttga
     2521 tattgacctc tatataacac ttgatgaaag ccaaaaaaga gaattgaaga atttactttt
     2581 agatatagtg gatgactata atttggtaaa tacaaaagat ttaatggctt ttattcgcct
     2641 taggggagcg gagtttggaa ttttaaatac gaatgatgta aaagatattg tttcaacaaa
     2701 ctctagcgcc tttagattat ggtttgaggg caattatcag tgtggatata gagcaagtta
     2761 tgcaaaggtt cttgatgctg aaacggggga aataaaatga caaacaaaga aaaagagtta
     2821 tttgctgaaa atgaggaatt aaaaaaagaa attaaggact taaaagagcg tattgaaaga
     2881 tacagagaaa tggaagttga attaagtaca acaatagatt tattgagagg agggattatt
     2941 gaataaataa aagccccctg acgaaagtcg aagggggttt ttattttggt ttgatgttgc
     3001 gattaatagc aatacaattg caataaacaa aatgatcctc tagagcatcc gcggagatag
     3061 gttatgcaag atttttatat ggaaatagca tgaaacaaaa atgaattaac tataattact
     3121 aaattcttat ttacagtcaa tcagattttt gattaaaagg agtagagaag tcatgagaac
     3181 ttatactttc gaccaggtag aaaaggcaat agagcagtta tatcctgatt ttactatcaa
     3241 tacaatagag atttcaggag aaggcaatga ctgtattgca tatgaaataa acagggattt
     3301 catttttaaa tttccaaagc attcaagagg atctactaat ctttttaatg aagtaaatat
     3361 actcaaaaga atccacaata aattacccct ccccattccg gaggtggttt ttacaggaat
     3421 gccatcagaa acgtaccaaa tgtctttcgc aggttttaca aaaattaaag gagtaccatt
     3481 gacacctctt ctactcaata atctgccgaa gcaatctcaa aatcaggcag ctaaggacct
     3541 ggcccgattt ctaagtgaac ttcacagcat aaacatctct ggattcaaaa gtaatctggt
     3601 attagatttt cgagagaaga taaatgaaga taataaaaaa atcaaaaagt tactatccag
     3661 ggaattaaag ggtccccaga tgaagatagt ggatgatttt tacagggata ttctagagaa
     3721 cgaaatctac ttcaaatact atccttgtct tattcataac gattttagca gtgatcatat
     3781 tttatttgat accgaaaaaa atactatttg tggaataatc gattttggag atgcagctat
     3841 ttctgatccc gacaatgatt ttataagttt gatggaagat gatgaagaat acggcatgga
     3901 atttgtatca aaaatattga accattacaa acataaggat ataccgacag ttttggaaaa
     3961 atataggatg aaagaaaaat actggtcgtt cgaaaagatt atctatggaa aggaatatgg
     4021 ttatatggat tggtatgaag agggattaaa tgaaatcaga agcattaaaa ttaaatagat
     4081 atttaatata aaggactgct aatgtactca tcaatagcaa tcctttttta gaatccggaa
     4141 tccgcgggga agggcgccgg cctaggacta gtttttatac tccccttgta ccaaaaggat
     4201 atgatttctt ggaggaaatt aaaaagttat ccaacagcag acagtccacg caggcgacgt
     4261 gcgaagcgga agtcgcaggt gtggactgat cttgctggct ggtgtggcaa tagccacgcc
     4321 agcacttaac cccccgtatc taacaggggg gtacaaatcg acaggaaaca gtcaaaaaaa
     4381 cattagaaaa tcctttggtt acaagggatt tacaaaattt cagcgtatgt caaatgggct
     4441 ttaaaagttg acatacgcct ttttgattgg agggattttt actgaatgaa caaacttggt
     4501 tacagcattt aaaagaaaaa cgcttggctt atggactatc tcaaaaccgt ttagctgttg
     4561 cgactggtat tacaaggcag tatctaagcg atattgaaac aggaaaagtc aagccatcag
     4621 aggatttaca gcagtccctt tgggaagctc tggaacgctt caatcccgac gctccccttg
     4681 aaatgctgtt tgattatgta aggattcgct ttccgacaac agacgtacag caggtggtcg
     4741 aaaacatctt acaactgaaa ctgtcctatt ttcttcatga ggactatggt ttctattctt
     4801 attcagagca ttatgcttta ggcgacatat tcgtcctttg acgcgtaaaa aaggcccaca
     4861 aaagtgggcc ttttttctcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4921 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5221 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5461 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccctcg
     5521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5581 ttgtttgcaa gcagcagatt acgtgcagaa aaaaaggatc tcaagaagat cctttgatct
     5641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5701 gattatcaaa aaggatcttc acctagatcc ttttaaatta gaaatgaagt tttaaatcaa
     5761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5881 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5941 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6001 gaagtggtcc gataaaccca gcgaaccatt tgaggtgata ggtaagatta taccgaggta
     6061 tgaaaacgag aattgaacct ttacagaatt actctatgaa gcgccatatt taaaagctac
     6121 caagaaaaaa agcggccaaa aaggcctgcc accaggccaa aaaggccgct tttttcgaag
     6181 aggatgaaga ggatgaggag gctgattgcc ttgaatatat tgacaatact gataagataa
     6241 tatatctttt atatagaaga ttttcgttcg tgaatacat
//
