| Plasmid Name | pET22b-SaCas9 | Promoter | T7 |
|---|---|---|---|
| Replicon | pBR322 | Copy Number | low |
| Competent Cell | DH5a | Culture Temp | 37°C |
| Forward Primer | T7：TAATACGACTCACTATAGGG | Reverse Primer | T7ter：TGCTAGTTATTGCTCAGCGG |
| 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.”
f1 ori 12..467
/direction=RIGHT
/note="f1 噬菌体复制起点；箭头指示(+)链合成方向"
AmpR promoter 494..598
/gene="bla"
AmpR 599..1459
/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
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
ori 1630..2218
/direction=RIGHT
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
bom 2404..2546
/note="basis of mobility region from pBR322"
rop complement(2648..2839)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
lacI complement(3648..4730)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="lac阻遏蛋白结合到lac操纵子上以抑制大肠杆菌中的转录。这种抑制可以通过添加乳糖或异丙基-beta-D-硫代半乳糖苷（IPTG）来解除。"
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
lacI promoter complement(4731..4808)
/gene="lacI"
T7 promoter 5117..5135
/note="1型人类免疫缺陷病毒RNA聚合酶启动子"
lac operator 5136..5160
/bound_moiety="lac repressor encoded by lacI"
/note="lac阻遏蛋白结合到lac操纵子上以抑制大肠杆菌中的转录。这种抑制可以通过添加乳糖或异丙基-β-D-硫代半乳糖苷（IPTG）来解除。"
RBS 5175..5197
/note="来自噬菌体T7基因10的高效核糖体结合位点（Olins和Rangwala，1989）"
sig_peptide 5205..5270
/gene="pelB (fragment)"
/product="leader peptide for secretion"
Cleavage site after base 5270"
6xHis 5337..5354
/codon_start=1
/product="6xHis affinity tag"
/translation="HHHHHH"
HA 5403..5429
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/translation="YPYDVPDYA"
SV40 NLS 5448..5468
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
CDS 5493..8648
/codon_start=1
/product="Cas9 endonuclease from the Staphylococcus aureus
Type II CRISPR/Cas system"
/note="在DNA中产生RNA引导的双链断裂"
/translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
IKKG"
nucleoplasmin NLS 8649..8696
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/translation="KRPAATKKAGQAKKKK"
6xHis 8697..8714
/codon_start=1
/product="6xHis affinity tag"
/translation="HHHHHH"
terminator 8784..8831
/note="1型大肠杆菌噬菌体T7 RNA聚合酶的转录终止子" 
