LOCUS       V003534                 4370 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003534
VERSION     V003534
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4370)
  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 4370)
  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 4370)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4370)
  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..4370
                     /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             1856..1945
                     /label="3xHA"
                     /note="three tandem HA epitope tags"
     misc_feature    1949..1978
                     /label="linker 2"
                     /note="linker 2"
     CDS             2157..2627
                     /gene="dhfrI"
                     /label="Dihydrofolate reductase type 1"
                     /note="Dihydrofolate reductase type 1 from Escherichia
                     coli. Accession#: P00382"
     primer_bind     complement(2858..2874)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(3309..3677)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3710..3819)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      3979..4367
                     /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 ggttctggca tcttttaccc
     1861 atacgatgtt cctgactatg cgggctatcc ctatgacgtc ccggactatg caggatccta
     1921 tccatatgac gttccagatt acgctgctgg ctctgcagga tctgctgcag gaggagctag
     1981 tttaaactct agatgcaagc aggatagacg gcatgcacga tttgtaataa cagagtgtct
     2041 tgtattttta gagaaagtct atttaataca agtgattata ttaattaacg gtaagcatca
     2101 gcgggtgaca aaacgagcat gcttactaat aaaatgttaa cctctgagga agaattgtga
     2161 aactatcact aatggtagct atatcgaaga atggagttat cgggaatggc cctgatattc
     2221 catggagtgc caaaggtgaa cagctcctgt ttaaagctat tacctataac caatggctgt
     2281 tggttggacg caagactttt ggatcaatgg gagcattacc caaccgaaag tatgcggtcg
     2341 taacacgttc aagttttaca tctgacaatg agaacgtatt gatctttcca tcaattaaag
     2401 atgctttaac caacctaaag aaaataacgg atcatgtcat tgtttcaggt ggtggggaga
     2461 tatacaaaag cctgatcgat caagtagata cactacatat atctacaata gacatcgagc
     2521 cggaaggtga tgtttacttt cctgaaatcc ccagcaattt taggccagtt tttacccaag
     2581 acttcgcctc taacataaat tatagttacc aaatctggca aaagggttaa caagtggcag
     2641 caacggattc gcaaacctgt cacgcctttt gtgccaaaag ccgcgccagg tttgcgatcc
     2701 gctgtgccag gcgttaggcg tcatatgaag atttcggtga tccctgagca ggtggcggaa
     2761 acattggatg ctgactgcag cctagtttaa gactttattg tcatagttta gatctttttt
     2821 gtccagtttg agactttttt gtgtttaaac agaattcact ggccgtcgtt ttacaacgtc
     2881 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2941 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3001 tgaatggcga atggggctcg atcccctcgc gagttggttc agctgctgcc tgaggctgga
     3061 cgacctcgcg gagttctacc ggcagtgcaa atccgtcggc atccaggaaa ccagcagcgg
     3121 ctatccgcgc atccatgccc ccgaactgca ggagtgggga ggcacgatgg ccgctttggt
     3181 cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga
     3241 tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa
     3301 aaaagtcatg gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc
     3361 tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg
     3421 cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg
     3481 atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg
     3541 ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc
     3601 gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc
     3661 ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct
     3721 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa
     3781 cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac
     3841 caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga
     3901 tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaacggccac
     3961 gatgcgtccg gcgtagagga tctgaagatc agcagttcaa cctgttgata gtacgtacta
     4021 agctctcatg tttcacgtac taagctctca tgtttaacgt actaagctct catgtttaac
     4081 gaactaaacc ctcatggcta acgtactaag ctctcatggc taacgtacta agctctcatg
     4141 tttcacgtac taagctctca tgtttgaaca ataaaattaa tataaatcag caacttaaat
     4201 agcctctaag gttttaagtt ttataagaaa aaaaagaata tataaggctt ttaaagcttt
     4261 taaggtttaa cggttgtgga caacaagcca gggatgtaac gcactgagaa gcccttagag
     4321 cctctcaaag caattttgag tgacacagga acacttaacg gctgacatgg
//
