Super PiggyBac Transposase vector (V012800)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V012800	Super PiggyBac Transposase	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

The Super PiggyBac transposase expression vector features a RNA polymerase II large subunit (Polr2A) promoter to provide robust expression of the transposase. The PiggyBac transposase coding sequence has been optimized for high expression, stability and integration activity in mammalian cells.

The PiggyBac (PB) transposon is a mobile genetic element that efficiently transposes between vectors andchromosomes via a "cut and paste" mechanism. During transposition, the PB transposase recognizes transposonspecific inverted terminal repeat sequences (ITRs) located on both ends of the transposon vector and efficiently moves the contents from the original sites and efficiently integrates them into TTAA chromosomal sites. The powerful activity of the piggyBac transposon system enables genes of interest between the two ITRs in the PB vector to be easily mobilized into target genomes. The Super PiggyBac transposase expression vector features a 5’ HS4 insulator to support robust transcription from the rPolr2A promoter.

The sequencing results indicated the XhoI site was destroyed by the insertion GTCGACGCCGCT.

Vector Name:: Super PiggyBac Transposase

Antibiotic Resistance:: Ampicillin

Length:: 8543 bp

Type:: PiggyBac

Replication origin:: ori

Source/Author:: SBI

Promoter:: rPolr2A

Growth Strain(s):: stbl3

Growth Temperature:: 37℃

Super PiggyBac Transposase 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

Sutrave G, Xu N, Tang TCY, Dolnikov A, Gloss B, Gottlieb DJ, Micklethwaite KP, Gowrishankar K. Characterizing piggyBat-a transposase for genetic modification of T cells. Mol Ther Methods Clin Dev. 2022 Mar 22;25:250-263.

Super PiggyBac Transposase vector Sequence

LOCUS       Exported                8543 bp DNA     circular SYN 26-AUG-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    Super PiggyBac Transposase
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8543)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8543)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8543)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3411..3422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..1993
                     /label=chicken 5'-HS4 beta-globin insulator (HS4)
                     /note="Insulators are DNA sequence elements that prevent 
                     inappropriate interactions between adjacent chromatin 
                     domains. One type of insulator establishes domains that 
                     separate enhancers and promoters to block their 
                     interaction, whereas a second type creates a barrier 
                     against the spread of heterochromatin."
     primer_bind     2449..2465
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2492..3197
                     /label=Polr2A promoter
                     /note="RNA polymerase II large subunit promoter"
     intron          3454..3550
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             3626..5407
                     /label=hyPBase
                     /note="Hyperactive version of piggyBac transposase (PBase) 
                     created by mutagenesis"
     polyA_signal    complement(5436..5570)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5687..5705)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5726..5742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5750..5766)
                     /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(5774..5804)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5819..5840)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6128..6716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6890..7747)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7748..7852)
                     /label=AmpR promoter
     rep_origin      7879..8334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8475..8491
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8501..8519
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"

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