pPK121 vector (V004079)

MSDS

Basic Vector Information

Vector Name:: pPK121

Antibiotic Resistance:: Ampicillin

Length:: 9915 bp

Type:: Expression vector

Replication origin:: ori

Source/Author:: Coy DL, Wagenbach M, Howard J.

pPK121 vector Vector Map

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Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pPK121 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V004079                 9915 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004079
VERSION     V004079
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9915)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Kinesin takes one 8-nm step for each ATP that it hydrolyzes
  JOURNAL   J. Biol. Chem. 274 (6), 3667-3671 (1999)
   PUBMED   9920916
REFERENCE   2  (bases 1 to 9915)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Mechanochemical Coupling Stoichiometry of Single Kinesin Motors
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 9915)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAR-1998) Physiology and Biophysics, University of
            Washington, Campus Box 357290, Seattle, WA 98195-7290, USA
REFERENCE   4  (bases 1 to 9915)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 9915)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Biol.
            Chem."; date: "1999"; volume: "274"; issue: "6"; pages: "3667-3671"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (23-MAR-1998) Physiology and Biophysics, University of Washington,
            Campus Box 357290, Seattle, WA 98195-7290, USA"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        494..598
                     /label="AmpR promoter"
     CDS             599..1456
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1630..2218
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2404..2546)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2651..2839)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3614..3635)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3651..4730)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4731..4808)
                     /label="lacI promoter"
     promoter        5117..5135
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5136..5160
                     /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)."
     RBS             5175..5197
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5205..8129
                     /gene="Khc"
                     /label="Kinesin heavy chain"
                     /note="Kinesin heavy chain from Drosophila melanogaster.
                     Accession#: P17210"
     CDS             8154..8177
                     /codon_start=1
                     /product="co-translated peptide"
                     /label="co-translated peptide"
                     /note="modeled on Schoner, B.E. et al. (1984) PNAS USA
                     81:5403-5407"
                     /protein_id="AAD13355.1"
                     /translation="MFPLEDD"
     CDS             8178..9701
                     /gene="Klc"
                     /label="Kinesin light chain"
                     /note="Kinesin light chain from Drosophila melanogaster.
                     Accession#: P46824"
     CDS             9702..9719
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             9720..9746
                     /label="9xHis"
                     /note="9xHis affinity tag"
     CDS             9759..9776
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      9843..9890
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"

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