pBC1 vector (V007018)

Price Information

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

Basic Vector Information

The pBC1 vector is a 21.6 kb vector designed to facilitate expression of recombinant proteins in the milk of transgenic animals. The pBC1 Milk Expression Vector Kit is intended for use in performing feasibility studies in mice, with the expectation that the user is interested in eventual large scale recombinant protein production using larger animals. Successful expression of recombinant protein in transgenic mice has generally been indicative of successful expression in larger animals such as goats or cows (Young et al., 1997). For feasibility studies, transgenic mice provide the added advantage of shorter generation times and faster evaluation than larger herd animals.

pBC1 vector Map

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.

pBC1 vector Sequence

LOCUS       V007018                21628 bp    DNA     circular SYN 13-JAN-2022
DEFINITION  Exported.
ACCESSION   V007018
VERSION     V007018
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 21628)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 21628)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..21628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(15009..15020)
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     promoter        15863..15967
                     /label="AmpR promoter"
     CDS             15968..16825
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      16998..17586
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(17772..17912)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(18017..18205)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             18772..19314
                     /gene="Nu1"
                     /label="Terminase small subunit"
                     /note="Terminase small subunit from Escherichia phage
                     lambda. Accession#: P03707"
     terminator      complement(20653..20747)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"