LOCUS       V003535                 4343 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003535
VERSION     V003535
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4343)
  AUTHORS   Zordan RE, Beliveau BJ, Trow JA, Craig NL, Cormack BP.
  TITLE     Transposon-based method for internal epitope tagging
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4343)
  AUTHORS   Zordan RE, Cormack BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JUN-2014) Molecular Biology and Genetics, Johns
            Hopkins School of Medicine, 725 N Wolfe St., Hunterian 609,
            Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 4343)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4343)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (04-JUN-2014) 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..4343
                     /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"
     repeat_region   1597..1795
                     /label="Tn7 left end"
                     /note="Tn7 left end"
     misc_feature    1820..1849
                     /label="linker 1"
                     /note="linker 1"
     CDS             1850..1921
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags"
     misc_feature    1922..1951
                     /label="linker 2"
                     /note="linker 2"
     CDS             2130..2600
                     /gene="dhfrI"
                     /label="Dihydrofolate reductase type 1"
                     /note="Dihydrofolate reductase type 1 from Escherichia
                     coli. Accession#: P00382"
     repeat_region   2754..2824
                     /label="Tn7 right end"
                     /note="Tn7 right end"
     primer_bind     complement(2831..2847)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(3282..3650)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3683..3792)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      3952..4340
                     /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 attacaagga
     1861 tgacgacgat aaggactata aggacgatga tgacaaggac tacaaagatg atgacgataa
     1921 aggctctgca ggatctgctg caggaggagc tagtttaaac tctagatgca agcaggatag
     1981 acggcatgca cgatttgtaa taacagagtg tcttgtattt ttagagaaag tctatttaat
     2041 acaagtgatt atattaatta acggtaagca tcagcgggtg acaaaacgag catgcttact
     2101 aataaaatgt taacctctga ggaagaattg tgaaactatc actaatggta gctatatcga
     2161 agaatggagt tatcgggaat ggccctgata ttccatggag tgccaaaggt gaacagctcc
     2221 tgtttaaagc tattacctat aaccaatggc tgttggttgg acgcaagact tttggatcaa
     2281 tgggagcatt acccaaccga aagtatgcgg tcgtaacacg ttcaagtttt acatctgaca
     2341 atgagaacgt attgatcttt ccatcaatta aagatgcttt aaccaaccta aagaaaataa
     2401 cggatcatgt cattgtttca ggtggtgggg agatatacaa aagcctgatc gatcaagtag
     2461 atacactaca tatatctaca atagacatcg agccggaagg tgatgtttac tttcctgaaa
     2521 tccccagcaa ttttaggcca gtttttaccc aagacttcgc ctctaacata aattatagtt
     2581 accaaatctg gcaaaagggt taacaagtgg cagcaacgga ttcgcaaacc tgtcacgcct
     2641 tttgtgccaa aagccgcgcc aggtttgcga tccgctgtgc caggcgttag gcgtcatatg
     2701 aagatttcgg tgatccctga gcaggtggcg gaaacattgg atgctgactg cagcctagtt
     2761 taagacttta ttgtcatagt ttagatcttt tttgtccagt ttgagacttt tttgtgttta
     2821 aacagaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     2881 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     2941 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggggc tcgatcccct
     3001 cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg
     3061 caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact
     3121 gcaggagtgg ggaggcacga tggccgcttt ggtcggtctt gccttgctcg tcggtgatgt
     3181 acttcaccag ctccgcgaag tcgctcttct tgatggagcg catggggacg tgcttggcaa
     3241 tcacgcgcac cccccggccg ttttagcggc taaaaaagtc atggctctgc cctcgggcgg
     3301 accacgccca tcatgacctt gccaagctcg tcctgcttct cttcgatctt cgccagcagg
     3361 gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag ccagagtttc
     3421 agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg gacgtgctca
     3481 tagtccacga cgcccgtgat tttgtagccc tggccgacgg ccagcaggta ggccgacagg
     3541 ctcatgccgg ccgccgccgc cttttcctca atcgctcttc gttcgtctgg aaggcagtac
     3601 accttgatag gtgggctgcc cttcctggtt ggcttggttt catcagccat ccgcttgccc
     3661 tcatctgtta cgccggcggt agccggccag cctcgcagag caggattccc gttgagcacc
     3721 gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca
     3781 cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg
     3841 caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc
     3901 ccgaagcagg gttatgcagc ggaaaacggc cacgatgcgt ccggcgtaga ggatctgaag
     3961 atcagcagtt caacctgttg atagtacgta ctaagctctc atgtttcacg tactaagctc
     4021 tcatgtttaa cgtactaagc tctcatgttt aacgaactaa accctcatgg ctaacgtact
     4081 aagctctcat ggctaacgta ctaagctctc atgtttcacg tactaagctc tcatgtttga
     4141 acaataaaat taatataaat cagcaactta aatagcctct aaggttttaa gttttataag
     4201 aaaaaaaaga atatataagg cttttaaagc ttttaaggtt taacggttgt ggacaacaag
     4261 ccagggatgt aacgcactga gaagccctta gagcctctca aagcaatttt gagtgacaca
     4321 ggaacactta acggctgaca tgg
//
