LOCUS       V003529                 4289 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003529
VERSION     V003529
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4289)
  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 4289)
  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 4289)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4289)
  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..4289
                     /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..1867
                     /label="6xHis"
                     /note="6xHis affinity tag"
     misc_feature    1868..1897
                     /label="linker 2"
                     /note="linker 2"
     CDS             2076..2546
                     /gene="dhfrI"
                     /label="Dihydrofolate reductase type 1"
                     /note="Dihydrofolate reductase type 1 from Escherichia
                     coli. Accession#: P00382"
     primer_bind     complement(2777..2793)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(3228..3596)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3629..3738)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      3898..4286
                     /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 ggttctggcc atcatcacca
     1861 tcaccacggc tctgcaggat ctgctgcagg aggagctagt ttaaactcta gatgcaagca
     1921 ggatagacgg catgcacgat ttgtaataac agagtgtctt gtatttttag agaaagtcta
     1981 tttaatacaa gtgattatat taattaacgg taagcatcag cgggtgacaa aacgagcatg
     2041 cttactaata aaatgttaac ctctgaggaa gaattgtgaa actatcacta atggtagcta
     2101 tatcgaagaa tggagttatc gggaatggcc ctgatattcc atggagtgcc aaaggtgaac
     2161 agctcctgtt taaagctatt acctataacc aatggctgtt ggttggacgc aagacttttg
     2221 gatcaatggg agcattaccc aaccgaaagt atgcggtcgt aacacgttca agttttacat
     2281 ctgacaatga gaacgtattg atctttccat caattaaaga tgctttaacc aacctaaaga
     2341 aaataacgga tcatgtcatt gtttcaggtg gtggggagat atacaaaagc ctgatcgatc
     2401 aagtagatac actacatata tctacaatag acatcgagcc ggaaggtgat gtttactttc
     2461 ctgaaatccc cagcaatttt aggccagttt ttacccaaga cttcgcctct aacataaatt
     2521 atagttacca aatctggcaa aagggttaac aagtggcagc aacggattcg caaacctgtc
     2581 acgccttttg tgccaaaagc cgcgccaggt ttgcgatccg ctgtgccagg cgttaggcgt
     2641 catatgaaga tttcggtgat ccctgagcag gtggcggaaa cattggatgc tgactgcagc
     2701 ctagtttaag actttattgt catagtttag atcttttttg tccagtttga gacttttttg
     2761 tgtttaaaca gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2821 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2881 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggggctcga
     2941 tcccctcgcg agttggttca gctgctgcct gaggctggac gacctcgcgg agttctaccg
     3001 gcagtgcaaa tccgtcggca tccaggaaac cagcagcggc tatccgcgca tccatgcccc
     3061 cgaactgcag gagtggggag gcacgatggc cgctttggtc ggtcttgcct tgctcgtcgg
     3121 tgatgtactt caccagctcc gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct
     3181 tggcaatcac gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg ctctgccctc
     3241 gggcggacca cgcccatcat gaccttgcca agctcgtcct gcttctcttc gatcttcgcc
     3301 agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag
     3361 agtttcagca ggccgcccag gcggcccagg tcgccattga tgcgggccag ctcgcggacg
     3421 tgctcatagt ccacgacgcc cgtgattttg tagccctggc cgacggccag caggtaggcc
     3481 gacaggctca tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg
     3541 cagtacacct tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc
     3601 ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg
     3661 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     3721 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     3781 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     3841 atgaccccga agcagggtta tgcagcggaa aacggccacg atgcgtccgg cgtagaggat
     3901 ctgaagatca gcagttcaac ctgttgatag tacgtactaa gctctcatgt ttcacgtact
     3961 aagctctcat gtttaacgta ctaagctctc atgtttaacg aactaaaccc tcatggctaa
     4021 cgtactaagc tctcatggct aacgtactaa gctctcatgt ttcacgtact aagctctcat
     4081 gtttgaacaa taaaattaat ataaatcagc aacttaaata gcctctaagg ttttaagttt
     4141 tataagaaaa aaaagaatat ataaggcttt taaagctttt aaggtttaac ggttgtggac
     4201 aacaagccag ggatgtaacg cactgagaag cccttagagc ctctcaaagc aattttgagt
     4261 gacacaggaa cacttaacgg ctgacatgg
//
