pAcGP67A vector (V005496) Gene synthesis in pAcGP67A backbone

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Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V005496	pAcGP67A	In stock, instant shipping

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

Basic Vector Information

Baculovirus transfer vector, pAcGP67A. Compatible with Spodoptera frugiperda Sf9 cells.

Vector Name:: pAcGP67A

Antibiotic Resistance:: Ampicillin

Length:: 9767 bp

Type:: Insect baculovirus expression vectors

Replication origin:: ori

Promoter:: polyhedrin

Growth Strain(s):: DH10B

Growth Temperature:: 37℃

pAcGP67A 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.

References

Afrin, L. B., Gulick, H., Vesely, J., Willingham, M., & Frankel, A. E. (1994). Expression of oligohistidine-tagged ricin B chain in Spodoptera frugiperda. Bioconjugate chemistry, 5(6), 539–546. https://doi.org/10.1021/bc00030a009

pAcGP67A vector Sequence

LOCUS       Exported                9767 bp DNA     circular SYN 24-OCT-2025
DEFINITION  Exported.
ACCESSION   V005496
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9767)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9767)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9767)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             76..579
                     /gene="PTP"
                     /label=Tyrosine-protein phosphatase
                     /note="Tyrosine-protein phosphatase from Autographa
                     californica nuclear polyhedrosis virus. Accession#: P24656"
     CDS             2662..3291
                     /gene="LEF-2"
                     /label=Primase-associated factor LEF-2
                     /note="Primase-associated factor LEF-2 from Autographa 
                     californica nuclear polyhedrosis virus. Accession#: P41418"
     misc_recomb     3170..3997
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     promoter        4001..4092
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     sig_peptide     4189..4248
                     /label=GP64 signal sequence
                     /note="signal sequence from the baculovirus envelope 
                     glycoprotein GP64"
     misc_recomb     4776..6231
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     primer_bind     complement(7148..7164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7172..7188)
                     /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)."
     promoter        complement(7196..7226)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7241..7262)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7550..8138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8312..9169)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9170..9274)
                     /label=AmpR promoter
     primer_bind     9748..9764
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"

.cls { stroke-width: 0px; fill: url(#c6); } pAcGP67A vector (V005496) Gene synthesis in pAcGP67A backbone