pMK4 vector (V004712)

Basic Vector Information

Vector Name:
pMK4
Antibiotic Resistance:
Ampicillin
Length:
5585 bp
Type:
Shuttle vector
Replication origin:
ori
Source/Author:
Zeigler DR.

pMK4 vector Vector Map

pMK45585 bp60012001800240030003600420048005400CAP binding sitelac promoterlac operatorM13 revM13 fwdAmpR promoterAmpRenterokinase siterepCChloramphenicol acetyltransferaseori

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 ℃.

pMK4 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V004712                 5585 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004712
VERSION     V004712
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5585)
  AUTHORS   Zeigler DR.
  TITLE     pMK3 and pMK4 revisited: nucleotide sequences of first generation
            Bacillus subtilis - E. coli shuttle vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5585)
  AUTHORS   Zeigler DR.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAR-2008) Bacillus Genetic Stock Center, The Ohio
            State University, 484 W 12th Ave, Columbus, OH 43210, USA
REFERENCE   3  (bases 1 to 5585)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5585)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-MAR-2008) Bacillus Genetic Stock Center, The Ohio State
            University, 484 W 12th Ave, Columbus, OH 43210, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(269..285)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        759..863
                     /label="AmpR promoter"
     CDS             864..1721
                     /label="AmpR"
                     /note="beta-lactamase"
     CDS             complement(2460..2474)
                     /label="enterokinase site"
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(2572..3507)
                     /codon_start=1
                     /gene="repC"
                     /product="replication initiation protein"
                     /label="repC"
                     /note="RepC; initiation of plasmid replication; derived
                     from Staphylococcus aureus plasmid pC194"
                     /protein_id="ACB41735.1"
                     /translation="MCYNMEKYTEKKQRNQVFQKFIKRHIGENQMDLVEDCNTFLSFVA
                     DKTLEKQKLYKANSCKNRFCPVCAWRKARKDALGLSLMMQYIKQQEKKEFIFLTLTTPN
                     VMSDELENEIKRYNNSFRKLIKRKKVGSVIKGYVRKLEITYNKKRDDYNPHFHVLIAVN
                     KSYFTDKRYYISQQEWLDLWRDVTGISEITQVQVQKIRQNNNKELYEMAKYSGKDSDYL
                     INQKVFDAFYKSLKGKQVLVYSGLFKEAKKKLKNGDLDYLKEIDPTEYIYQIFYIWKQK
                     EYLASELYDLTEQEKREINHKMIDEIEEEQ"
     gene            complement(2572..3507)
                     /gene="repC"
                     /label="repC"
     CDS             3965..4612
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     rep_origin      4815..5403
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"

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