LOCUS       40924_45038        4466 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18R6K-mini-Tn7T-Km, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4466)
  AUTHORS   Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     A Tn7-based broad-range bacterial cloning and expression system
  JOURNAL   Nat. Methods 2 (6), 443-448 (2005)
  PUBMED    15908923
REFERENCE   2  (bases 1 to 4466)
  AUTHORS   Choi K-H., Gaynor JB, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2004) Microbiology, Immunology and Pathology, 
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 4466)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4466)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2005"; volume: "2"; issue: "6"; pages: "443-448"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAY-2004) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   382..580
                     /note="Tn7R; right end of Tn7"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      826..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    966..1013
                     /label=FRT, Flp recombinase target site
                     /note="FRT, Flp recombinase target site"
     protein_bind    complement(966..1013)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             1163..1975
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    complement(2088..2135)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    2154..2231
                     /label=MCS, multiple cloning site
                     /note="MCS, multiple cloning site"
     mobile_element  complement(2240..2405)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2641..2997
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3409..4266)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4267..4371)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
      361 tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
      421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
      481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
      541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacggtatc
      601 gataagctag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      721 aatccaagct agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
      781 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      901 agtaggacaa atccgccgct aggagcttgc ggcccggacg atcgaattag cttcaaaagc
      961 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac ttcaagatcc
     1021 cctgattccc tttgtcaaca gcaatggatg ggaaagccac gttgtgtctc aaaatctctg
     1081 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1141 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctccaggcc
     1201 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggcac gcgataatgt
     1261 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     1321 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1381 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1441 tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata
     1501 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     1561 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     1621 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     1681 gcctgttgaa caagtctgga aagaaatgca taagctcttg ccattctcac cggattcagt
     1741 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1801 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1861 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1921 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1981 agaattggtt aattggttgt aacactggca gagcattacg ctgacttccc atctagaatt
     2041 aattccattg agtaagtttt taagcacatc agcttcaaaa gcgctctgaa gttcctatac
     2101 tttctagaga ataggaactt cggaatagga acttcaagat ccccaattcg atcatgcatg
     2161 agctcactag tggatccccc gggctgcagg aattcctcga gaagcttggg cccggtacct
     2221 cgcgaaggcc ttgcaggcca accagataag tgaaatctag ttccaaacta ttttgtcatt
     2281 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     2341 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     2401 ccacagcggg gcatttttct tcctgttatg tttgggcgct gcattaatga atcggccaac
     2461 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2581 tatccacaga atcaggggat aacgcaggaa agaacatggc ggcagttcaa cctgttgata
     2641 gtacgtacta agctctcatg tttcacgtac taagctctca agtttaacgt actaagctct
     2701 catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc taacgtacta
     2761 agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa tataaatcag
     2821 caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata tataaggctt
     2881 ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac gcactgagaa
     2941 gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg gctgacaacc
     3001 ggctggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3061 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3121 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3181 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3241 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3301 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3361 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3421 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3481 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3541 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3601 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3661 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3721 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3781 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3841 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3901 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3961 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4021 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4081 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4141 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4201 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4261 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4321 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4381 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4441 taggcgtatc acgaggccct ttcgtc
//
