pMK4 vector (V004712)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V004712	pMK4	In stock, 1 week for quality controls	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

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

View Map in NovoBuilder

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

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  JOURNAL   Unpublished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"