pFKM2 vector (V006249)

Basic Vector Information

Vector Name:
pFKM2
Antibiotic Resistance:
Ampicillin
Length:
4154 bp
Type:
Kanamycin resistance FRT vector
Replication origin:
ori
Source/Author:
Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer HP.

pFKM2 vector Map

pFKM24154 bp60012001800240030003600M13 fwdMCSFRTNeoR/KanRFRT; Flp recombinase target siteMCSM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

pFKM2 vector Sequence

LOCUS       40924_20081        4154 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Kanamycin resistance FRT vector pFKM2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4154)
  AUTHORS   Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer 
            HP.
  TITLE     Genetic tools for select-agent-compliant manipulation of 
            Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 74 (4), 1064-1075 (2008)
  PUBMED    18156318
REFERENCE   2  (bases 1 to 4154)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Genetic tools for Pseudomonas and its related bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4154)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-SEP-2007) Microbiology, Immunology and Pathology, 
            Colorado State University, 1682 Campus Delivery, Ft. Collins, CO 
            80523-1682, USA
REFERENCE   4  (bases 1 to 4154)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4154)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "4"; pages:
            "1064-1075"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (21-SEP-2007) Microbiology, Immunology and Pathology, Colorado State
            University, 1682 Campus Delivery, Ft. Collins, CO 80523-1682, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(401..457)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    466..513
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(623..1414)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    1800..1847
                     /note="FRT; Flp recombinase target site"
     protein_bind    1800..1847
                     /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)."
     misc_feature    1864..1920
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1933..1949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1957..1973)
                     /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)."
     promoter        complement(1981..2011)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2026..2047)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2335..2923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3097..3954)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3955..4059)
                     /label=AmpR promoter

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