| Plasmid Name | pX462 V2.0 | Alternative Names | 等效质粒Plasmid#62987;pSpCas9n(BB)-2A-Puro(PX462)V2.0 |
|---|---|---|---|
| Promoter | U6 | Replicon | pUC |
| Copy Number | high | Competent Cell | Stbl3 |
| Culture Temp | 37°C | Forward Primer | U6-F：GAGGGCCTATTTCCCATGA |
| Reverse Primer | BGH：TAGAAGGCACAGTCGAGG | Storage Temp | 37°C |
1.For Research Use Only
All products available through the MiaoLing Plasmid Platform are intended for research use only. They are not intended for clinical trials or for use in humans, including oral administration, injection, or therapeutic use in vivo or ex vivo.
2.Plasmid Sequence and Experimental Performance
Plasmid products are provided and sold based on the corresponding reference/template plasmid. MiaoLing guarantees the accuracy of the specified gene sequence based on our sequencing results, but does not guarantee downstream experimental performance, including gene expression, transfection efficiency, protein expression, or other experimental outcomes.
3.Sequence Verification of Certain Vector Products
For certain vector products, even the original manufacturer may not have fully verified the complete plasmid sequence. If the sequence of the key functional regions has been confirmed to be correct, the product will be considered a correct and usable vector for research purposes.
4.After-Sales Period for Small-Pack Plasmid Products
The after-sales service period for small-pack plasmid products is three months from the date of receipt. Customers are advised to transform and process the plasmid promptly after receipt. After the three-month period, a new purchase may be required for further support or replacement.
5.Research-Related Risks and Limitation of Liability
Scientific research is inherently exploratory and involves a significant degree of uncertainty. To the maximum extent permitted by applicable law, MiaoLing shall not be liable for any additional economic losses, indirect losses, or other liabilities arising from the use of our products or from research outcomes.
6.Citation in Chinese Publications
For publications in Chinese, please cite the product as follows:
7.Product Name (Product Item Number) was provided by the MiaoLing Plasmid Platform.
Citation in English Publications
For publications in English, please cite the product as follows:
“Product Name (Product Item Number) was obtained from MiaoLing Plasmid, China.”
U6 promoter 1..241
/note="人类U6 snRNA的RNA聚合酶III启动子"
misc_RNA 268..343
/note="CRISPR/Cas9系统指南RNA骨架"
chicken beta-actin promoter 727..1004
CDS 1254..1319
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
Cleavage site after base 1319"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
SV40 NLS 1326..1346
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/translation="PKKKRKV"
CDS 1371..5471
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/note="由于RuvC样催化域中的D10突变，产生RNA引导的DNA单链切口。"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
nucleoplasmin NLS 5472..5519
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/translation="KRPAATKKAGQAKKKK"
T2A 5535..5588
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/note="真核核糖体无法在甘氨酸和脯氨酸残基之间插入肽键，产生分离的多肽。"
Cleavage site after base 5585"
/translation="EGRGSLLTCGDVEENPGP"
PuroR 5589..6188
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="赋予对普鲁霉素的抗性"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
bGH poly(A) signal 6222..6429
/note="牛生长激素polyA信号"
repeat_region 6438..6578
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
f1 ori 6653..7108
/direction=RIGHT
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
AmpR promoter 7390..7494
/gene="bla"
AmpR 7495..8355
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="赋予对氨苄青霉素、卡那霉素和相关抗生素的抗性。"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
ori 8526..9114
/direction=RIGHT
/note="高拷贝数 ColE1/pMB1/pBR322/pUC 复制起点" 
