Basic Vector Information
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).
- Vector Name:
- Mega PiggyBac Transposase
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8543 bp
- Type:
- PiggyBac
- Copy Number:
- High copy number
- Promoter:
- rPolr2A
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
Mega PiggyBac Transposase vector Map
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 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"
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