Mega PiggyBac Transposase vector (Cat. No.: V018124)

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Basic Information

Note: Mega-PiggyBac is a hyperactive synthetic variant of the PiggyBac transposase, engineered through fine-tuning the ProGen2 protein language model (pLLM) using over 13,000 bioprospected PiggyBac sequences. Derived from targeted optimization of the laboratory-evolved hyperactive PiggyBac (HyPB), it stands out for its superior transposition efficiency—exhibiting both significantly higher excision activity and nontargeted integration activity compared to HyPB in HEK293T cells. Notably, Mega-PiggyBac (designated as seq3277) demonstrates excellent compatibility with advanced genome-editing systems, such as the FiCAT targeted integration platform, enabling a twofold improvement in site-specific insertion at loci like AAVS1-3, TTR, and PCSK9. Its robust performance extends to critical therapeutic contexts, including functional activity in primary T cells, making it a promising tool for gene therapy and synthetic biology applications that require efficient and precise large DNA fragment integration. This plasmid shares the same backbone as the Super PiggyBac Transposase vector (V012800).

Name:: Mega PiggyBac Transposase

Antibiotic Resistance:: Ampicillin

Length:: 8543 bp

Type:: PiggyBac

Copy Number:: High copy number

Promoter:: rPolr2A

Growth Strain(s):: Top10

Growth Temperature:: 37℃

$ 198.8

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

Resources

Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add sterile water to dissolve the DNA: add 20 μl for 5 μg plasmid, and 100 μl for 100 μg plasmid.

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

Ivančić D, Agudelo A, Lindstrom-Vautrin J, Jaraba-Wallace J, Gallo M, Das R, Ragel A, Herrero-Vicente J, Higueras I, Billeci F, Sanvicente-García M, Petazzi P, Ferruz N, Sánchez-Mejías A, Güell M. Discovery and protein language model-guided design of hyperactive transposases. Nat Biotechnol. 2025 Oct 2. doi: 10.1038/s41587-025-02816-4. Epub ahead of print. PMID: 41039042.

Mega PiggyBac Transposase vector (Cat. No.: V018124) Sequence

LOCUS       Exported                8543 bp DNA     circular SYN 14-NOV-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    Mega 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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8543)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; 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"
     promoter        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
                     /codon_start=1
                     /product="Mega-PiggyBac"
                     /label=Mega-PiggyBac
                     /note="Mega-PiggyBac"
                     /translation="MGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEE
                     AFIDEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKPT
                     RRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESM
                     TSATFRDTNEDEIRAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIR
                     CLRMDDKSIRPTLRESDVFTPVRKIWDIFINQCIQNYTPGAHLTIDEQLLGFRGRCPFR
                     VYIPNKPSKYGIKIVMICDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVRGSC
                     RNITCDNWFTSIPLAKNLLQEPYKLTLVGTVRSNKREIPEELKNSRSRPVGTSMFCFDG
                     PLTLVSYKPKPAKMVYLLSSCDEDAIINETTGKPEMIMYYNQTKGGVDTLDQMCSLMSC
                     SRKTNRWPMALFYGMINIACINSYIIYCHNVSSKGEKVQSRKKFMKNLYKGLTSSFMRK
                     RLEAPTLKRYLRDNISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKASASCKK
                     CKKVICREHNIDMCQSCF"
     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"