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pBlueBacHis2 A vector (V010672)

Price Information

Cat No. Plasmid Name Availability Add to cart
V010672 pBlueBacHis2 A In stock, 1 week for quality controls

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

Basic Vector Information

pBlueBacHis2 A (4863 bp), pBlueBacHis2 B (4853 bp), and pBlueBacHis2 C (4861 bp) are baculovirus transfer vectors that contain the Xpress N-terminal tag for expression and purification of recombinant fusion proteins. The pBlueBacHis2 vector is provided in three different versions for simplified cloning of your gene in-frame with the Xpress tag. The vectors contain sequences homologous to the lacZ gene and ORF1629 sequences in Bac-N-Blue linear DNA, allowing production of blue, recombinant plaques. Expression of your recombinant fusion protein is driven by the polyhedrin promoter.

Vector Name:
pBlueBacHis2 A
Antibiotic Resistance:
Ampicillin
Length:
4863 bp
Type:
Insect Cell Expression Vectors
Replication origin:
ori
Copy Number:
High copy number
Promoter:
polyhedrin
Cloning Method:
Enzyme digestion and ligation
5' Primer:
polyhedrin fwd
Fusion Tag:
His

pBlueBacHis2 A vector Map

Copy Sequence View Map
pBlueBacHis2 A4863 bp6001200180024003000360042004800polyhedrin promoter6xHisT7 tag (gene 10 leader)Xpress(TM) tagcontains part of ORF1629AmpR promoterAmpRori

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.

pBlueBacHis2 A vector Sequence

LOCUS       40924_6711        4863 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4863)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4863)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..94
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             110..127
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             131..163
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             167..190
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     misc_recomb     267..972
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     promoter        1548..1652
                     /label=AmpR promoter
     CDS             1653..2510
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2684..3272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"