pLMF03 vector (V004971)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V004971	pLMF03	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:: pLMF03

Antibiotic Resistance:: Ampicillin

Length:: 6733 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Liu M, Wang M.

pLMF03 vector Map

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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.

pLMF03 vector Sequence

LOCUS       V004971                 6733 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004971
VERSION     V004971
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6733)
  AUTHORS   Liu M, Wang M.
  TITLE     Development of a plasmid-based methods for expression and antisense
            gene knockdown in Riemerella anatipestifer and application
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6733)
  AUTHORS   Liu M, Wang M.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAR-2016) Institute of Preventive Veterinary Medicine,
            Sichuan Agricultural University, No. 211, Huimin Road, Chengdu,
            Sichuan 611130, China
REFERENCE   3  (bases 1 to 6733)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6733)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (26-MAR-2016) Institute of Preventive Veterinary Medicine, Sichuan
            Agricultural University, No. 211, Huimin Road, Chengdu, Sichuan
            611130, China"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    290..311
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        326..356
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    364..380
                     /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     388..404
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             2690..2707
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             complement(3379..4341)
                     /gene="cfxA"
                     /label="Beta-lactamase"
                     /note="Beta-lactamase from Phocaeicola vulgatus.
                     Accession#: P30899"
     primer_bind     complement(4521..4537)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        5011..5115
                     /label="AmpR promoter"
     CDS             5116..5973
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      6147..6733
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"