LOCUS       Exported                4146 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pFKm4, complete sequence.
ACCESSION   JX504717
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4146)
  AUTHORS   Kvitko BH, Bruckbauer S, Prucha J, McMillan I, Breland EJ, Lehman S,
            Mladinich K, Choi KH, Karkhoff-Schweizer R, Schweizer HP.
  TITLE     A simple method for construction of pir+ Enterobacterial hosts for 
            maintenance of R6K replicon plasmids
  JOURNAL   BMC Res Notes 5, 157 (2012)
  PUBMED    22433797
REFERENCE   2  (bases 1 to 4146)
  AUTHORS   Kvitko BH, Goodyear A, Propst KL, Dow SW, Schweizer HP.
  TITLE     Burkholderia pseudomallei Known Siderophores and Hemin Uptake Are 
            Dispensable for Lethal Murine Melioidosis
  JOURNAL   PLoS Negl Trop Dis 6 (6), E1715 (2012)
  PUBMED    22745846
REFERENCE   3  (bases 1 to 4146)
  AUTHORS   Kvitko BH, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-AUG-2012) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   4  (bases 1 to 4146)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4146
                     /organism="Plasmid vector pFKm4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1245209"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     466..513
                     /label=FRT
                     /note="FRT"
     protein_bind    466..513
                     /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(612..1406)
                     /codon_start=1
                     /gene="nptII"
                     /product="NptII"
                     /EC_number="2.7.1.95"
                     /label=nptII
                     /note="neomycin phosphotransferase; kanamycin resistance"
                     /protein_id="AFV53145.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(612..1406)
                     /gene="nptII"
                     /label=nptII
     misc_recomb     1792..1839
                     /label=FRT
                     /note="FRT"
     protein_bind    1792..1839
                     /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)."
     primer_bind     complement(1925..1941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1949..1965
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(1973..2003)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2018..2039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2245..2918
                     /label=pUC origin
                     /note="pUC origin"
     rep_origin      complement(2327..2915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3086..3946)
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /EC_number="3.5.2.6"
                     /label=bla
                     /note="beta-lactamase; ampicillin resistance"
                     /protein_id="AFV53146.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3086..3946)
                     /gene="bla"
                     /label=bla
     promoter        complement(3947..4051)
                     /gene="bla"
                     /label=AmpR promoter