LOCUS       V003530                 4316 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003530
VERSION     V003530
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4316)
  AUTHORS   Zordan RE, Beliveau BJ, Trow JA, Craig NL, Cormack BP.
  TITLE     Avoiding the ends: internal epitope tagging of proteins using
            transposon Tn7
  JOURNAL   Genetics 200 (1), 47-58 (2015)
   PUBMED   25745023
REFERENCE   2  (bases 1 to 4316)
  AUTHORS   Zordan RE, Cormack BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2015) Molecular Biology and Genetics, Johns
            Hopkins School of Medicine, 725 N Wolfe St., Hunterian 609,
            Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 4316)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4316)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics";
            date: "2015"; volume: "200"; issue: "1"; pages: "47-58"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (24-JAN-2015) Molecular Biology and Genetics, Johns Hopkins School
            of Medicine, 725 N Wolfe St., Hunterian 609, Baltimore, MD 21205,
            USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             361..1152
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     protein_bind    1430..1451
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1466..1496
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1504..1520
                     /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)."
     primer_bind     1528..1544
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    1820..1849
                     /label="linker 1"
                     /note="linker 1"
     CDS             1850..1894
                     /label="AviTag(TM)"
                     /note="peptide tag that allows for enzymatic biotinylation"
     misc_feature    1895..1924
                     /label="linker 2"
                     /note="linker 2"
     CDS             2103..2573
                     /gene="dhfrI"
                     /label="Dihydrofolate reductase type 1"
                     /note="Dihydrofolate reductase type 1 from Escherichia
                     coli. Accession#: P00382"
     primer_bind     complement(2804..2820)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(3255..3623)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3656..3765)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      3925..4313
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gaattagctt cacgctgccg caagcactca gggcgcaagg gctgctaaag gaagcggaac
       61 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
      121 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
      181 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
      241 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
      301 aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc
      361 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
      421 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
      481 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
      541 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
      601 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
      661 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
      721 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
      781 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
      841 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
      901 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
      961 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     1021 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     1081 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     1141 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     1201 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     1261 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     1321 cccacccccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     1381 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     1441 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     1501 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     1561 ttgcatgcct gcaggtcgac tctagctaga ggatcctgtg gccggccaat aaagtcttaa
     1621 actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag aatagttgta
     1681 aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag
     1741 caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggggtcg
     1801 acggcgcgcc ggccggcctg gatccgctgg ctccgctgct ggttctggcg gtttgaacga
     1861 tatttttgaa gctcaaaaaa ttgaatggca tgaaggctct gcaggatctg ctgcaggatc
     1921 tggaagttta aactctagat gcaagcagga tagacggcat gcacgatttg taataacaga
     1981 gtgtcttgta tttttagaga aagtctattt aatacaagtg attatattaa ttaacggtaa
     2041 gcatcagcgg gtgacaaaac gagcatgctt actaataaaa tgttaacctc tgaggaagaa
     2101 ttgtgaaact atcactaatg gtagctatat cgaagaatgg agttatcggg aatggccctg
     2161 atattccatg gagtgccaaa ggtgaacagc tcctgtttaa agctattacc tataaccaat
     2221 ggctgttggt tggacgcaag acttttggat caatgggagc attacccaac cgaaagtatg
     2281 cggtcgtaac acgttcaagt tttacatctg acaatgagaa cgtattgatc tttccatcaa
     2341 ttaaagatgc tttaaccaac ctaaagaaaa taacggatca tgtcattgtt tcaggtggtg
     2401 gggagatata caaaagcctg atcgatcaag tagatacact acatatatct acaatagaca
     2461 tcgagccgga aggtgatgtt tactttcctg aaatccccag caattttagg ccagttttta
     2521 cccaagactt cgcctctaac ataaattata gttaccaaat ctggcaaaag ggttaacaag
     2581 tggcagcaac ggattcgcaa acctgtcacg ccttttgtgc caaaagccgc gccaggtttg
     2641 cgatccgctg tgccaggcgt taggcgtcat atgaagattt cggtgatccc tgagcaggtg
     2701 gcggaaacat tggatgctga ctgcagccta gtttaagact ttattgtcat agtttagatc
     2761 ttttttgtcc agtttgagac ttttttgtgt ttaaacagaa ttcactggcc gtcgttttac
     2821 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2881 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2941 gcagcctgaa tggcgaatgg ggctcgatcc cctcgcgagt tggttcagct gctgcctgag
     3001 gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag
     3061 cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca cgatggccgc
     3121 tttggtcggt cttgccttgc tcgtcggtga tgtacttcac cagctccgcg aagtcgctct
     3181 tcttgatgga gcgcatgggg acgtgcttgg caatcacgcg caccccccgg ccgttttagc
     3241 ggctaaaaaa gtcatggctc tgccctcggg cggaccacgc ccatcatgac cttgccaagc
     3301 tcgtcctgct tctcttcgat cttcgccagc agggcgagga tcgtggcatc accgaaccgc
     3361 gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg gcccaggtcg
     3421 ccattgatgc gggccagctc gcggacgtgc tcatagtcca cgacgcccgt gattttgtag
     3481 ccctggccga cggccagcag gtaggccgac aggctcatgc cggccgccgc cgccttttcc
     3541 tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga taggtgggct gcccttcctg
     3601 gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc
     3661 cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag
     3721 aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg
     3781 atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag
     3841 gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaac
     3901 ggccacgatg cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac
     3961 gtactaagct ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg
     4021 tttaacgaac taaaccctca tggctaacgt actaagctct catggctaac gtactaagct
     4081 ctcatgtttc acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac
     4141 ttaaatagcc tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa
     4201 agcttttaag gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc
     4261 ttagagcctc tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgg
//
