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pBAC-5 vector (V011670)

Price Information

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V011670 pBAC-5 In stock, instant shipping

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

Basic Vector Information

pBAC-5 is a baculovirus transfer plasmid with the gp64 promoter, encoding an N-terminal 6xHis-thrombin-S-Tag-enterokinase cassette.

Vector Name:
pBAC-5
Antibiotic Resistance:
Ampicillin
Length:
5517 bp
Type:
Insect Cell Vectors
Replication origin:
ori
Source/Author:
Novagen (EMD Millipore)
Copy Number:
High copy number
Promoter:
gp64
Growth Strain(s):
stbl3
Growth Temperature:
37℃

pBAC-5 vector Map

Copy Sequence View Map
pBAC-55517 bp60012001800240030003600420048005400contains ORF603 and part of lef2gp64 promoterATG6xHisthrombin siteS-Tagenterokinase siteMCS6xHiscontains part of ORF1629oriAmpRAmpR promoterf1 ori

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.

References

  • Xu X, Zhou X, Nan H, Zhao Y, Bai Y, Ou Y, Chen H. Aggregation of AcMNPV LEF-10 and Its Impact on Viral Late Gene Expression. PLoS One. 2016 May 6;11(5):e0154835.

pBAC-5 vector Sequence

LOCUS       pBAC-5.        5517 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Baculovirus transfer plasmid with the gp64 promoter, encoding an 
            N-terminal 6xHis-thrombin-S-Tag-enterokinase cassette.
ACCESSION   .
VERSION     .
KEYWORDS    pBAC-5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5517)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5517)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     329..1156
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     promoter        1166..1369
                     /label=gp64 promoter
                     /note="promoter for the baculovirus gp64 glycoprotein gene"
     CDS             1372..1374
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             1378..1395
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             1405..1422
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             1426..1470
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             1486..1500
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     misc_feature    1502..1566
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1567..1584
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_recomb     1621..2978
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     rep_origin      complement(3241..3829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4003..4860)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4861..4965)
                     /label=AmpR promoter
     rep_origin      4992..5447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"