LOCUS       Exported                6103 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pR6KT-miniTn7T-P1eGFP-FK, complete sequence.
ACCESSION   KC577243
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6103)
  AUTHORS   Kvitko B, Bruckbauer S.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-FEB-2013) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   2  (bases 1 to 6103)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6103
                     /organism="Cloning vector pR6KT-miniTn7T-P1eGFP-FK"
                     /mol_type="other DNA"
                     /db_xref="taxon:1332675"
     protein_bind    18..65
                     /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             complement(172..966)
                     /codon_start=1
                     /gene="nptII"
                     /product="NptII"
                     /label=nptII
                     /note="kanamycin resistance protein"
                     /protein_id="AGL73306.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(172..966)
                     /gene="nptII"
                     /label=nptII
     protein_bind    1352..1399
                     /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             complement(1481..2278)
                     /codon_start=1
                     /gene="gfp"
                     /product="Gfp"
                     /label=gfp
                     /note="green fluorescent protein"
                     /protein_id="AGL73305.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICATGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFKGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGRTQISSSSFEFCSRRYRGPGIHRI"
     gene            complement(1481..2278)
                     /gene="gfp"
                     /label=gfp
     promoter        complement(2604..2632)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     repeat_region   2708..2873
                     /label=Tn7L
                     /note="Tn7L"
     mobile_element  2708..2873
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3177..3285
                     /note="incP origin of transfer"
     rep_origin      3316..3697
                     /label=oriV R6K
                     /note="oriV R6K"
     rep_origin      3336..3692
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(4101..4961)
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="ampicillin resistance protein"
                     /protein_id="AGL73307.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4101..4961)
                     /gene="bla"
                     /label=bla
     promoter        complement(4962..5066)
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   5543..5741
                     /label=Tn7R
                     /note="Tn7R"
     primer_bind     5746..5762
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5790..5884
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      5987..6073
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"