我们所展示的质粒图谱主要是从文献和开放数据库中收集而来，主要是为了方便研究工作，其中一小部分质粒进行了质量控制，可供科学家使用。
所有的产品都严格仅供科学研究使用，不能应用于临床试验，包括人体摄入、注射或外用的药理学用途。
确保质粒的关键元件正确，但是我们并不能保证实验结果。页面展示的图谱序列为理论序列，可能与测序结果不一致，请自行比对后确定是否满足要求。(如果测过序，本页面一般会提供下载)
开放数据库中的大多数载体序列都没有被完全测序。如果实际序列与参考序列的相似度超过99%，则将其视为正确。
由于科学研究是在探索未知，具有很大的不确定性，在任何情况下，我们都不承担超出质粒本身的额外经济损失或责任。
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).
- 载体名称:
- Mega PiggyBac Transposase
- 载体抗性:
- Ampicillin
- 载体长度:
- 8543 bp
- 载体类型:
- PiggyBac
- 拷贝数:
- High copy number
- 启动子:
- rPolr2A
- 感受态:
- DH10B
- 培养温度:
- 37℃
Mega PiggyBac Transposase 载体图谱
质粒操作方法
1. 发货形式：质粒干粉(常温运输，存于-20度，请务必先转化提质粒后使用)
2. 收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O溶解质粒;（质粒复测的浓度有时候与标称值差距较大，这可能是因为冻干质粒在管中的位置、复溶效率、测量偏差以及管壁的吸附导致，因此建议先转化提质粒后再使用）
3. 取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；
4. 加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min (30℃培养1-1.5小时);
5. 6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上;
6. 将平板倒置37℃培养14h，如果要求是30℃则培养20h; (菌落过多则将质粒稀释后再转化；没有菌落则加入10μl质粒转化；建议不要直接转表达感受态， 要先转克隆感受态，重提质粒后再导入表达感受态);
7. 挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
Mega PiggyBac Transposase 载体序列
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"