产品声明
- 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. - 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. - 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. - 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. - 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. - Citation in Chinese Publications
For publications in Chinese, please cite the product as follows:
产品名称（产品货号）由淼灵质粒平台提供。 - 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.”
质粒元件
f1 ori complement(14..469)
/direction=LEFT
/note="f1噬菌体复制起点；箭头表示（+）链合成的方向"
primer_bind 611..627
/note="常见的测序引物，多种相似变体之一"
TRE3G promoter 1268..1646
/note="可通过Tet-On（R）3G结合激活的第三代Tet反应启动子"
tet operator 1276..1294
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tetR和tetA基因的细菌操作员O2"
tet operator 1312..1330
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tetR和tetA基因的细菌操作员O2"
tet operator 1348..1366
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tetR和tetA基因的细菌操作员O2"
tet operator 1384..1402
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tetR和tetA基因的细菌操作员O2"
tet operator 1420..1438
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tetR和tetA基因的细菌操作员O2"
tet operator 1456..1474
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tetR和tetA基因的细菌操作员O2"
tet operator 1492..1510
/gene="tetO"
/bound_moiety="tetracycline repressor TetR"
/note="tetR和tetA基因的细菌操作员O2"
regulatory 1673..1682
/regulatory_class="other"
/note="脊椎动物强启动翻译的共识序列（Kozak，1987）"
CDS 1679..5782
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/note="RNA引导的DNA结合蛋白，由于D10A、D839A、H840A和N863A突变而缺乏内切酶活性（Mali等人，2013）"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
SV40 NLS 5795..5815
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
CDS 5840..5989
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
SV40 NLS 6014..6034
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
CDS 6464..6820
/codon_start=1
/product="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
/translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
EDFSSIADMDFSALL"
CDS 6839..7408
/codon_start=1
/product="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
/translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
DTFLNDECLLHAMHISTGLSIFDTSLF"
HSV TK poly(A) signal 7507..7555
/note="单纯疱疹病毒胸苷激酶多聚腺苷酸化信号（Cole和Stacy，1985）"
EF-1-alpha core promoter 7786..7997
/note="人延长因子EF-1α的核心启动子"
5' LTR (truncated) 8010..8278
/note="人T细胞白血病病毒1型截短5'长末端重复序列"
CDS 8312..9055
/codon_start=1
/product="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
/translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
T2A 9056..9109
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/note="真核核糖体不能在Gly和Pro残基之间插入肽键，从而产生单独的多肽。"
Cleavage site after base 9106"
/translation="EGRGSLLTCGDVEENPGP"
HygR 9131..10150
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/note="赋予对潮霉素的抗性"
/translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
SV40 poly(A) signal 10261..10382
/note="SV40聚腺苷酸化信号"
primer_bind complement(11670..11686)
/note="常见的测序引物，多种相似变体之一"
lac operator 11694..11710
/bound_moiety="lac repressor encoded by lacI"
/note="lac阻遏物与lac操作员结合，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-硫代吡喃半乳糖苷（IPTG）来缓解。"
lac promoter complement(11718..11748)
/note="大肠杆菌lac操纵子的启动子"
CAP binding site 11763..11784
/bound_moiety="E. coli catabolite activator protein"
/note="CAP结合在cAMP存在下激活转录。"
ori complement(12072..12660)
/direction=LEFT
/note="高拷贝数ColE1/pMB1/pBR322/pUC复制起点"
AmpR complement(12831..13691)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="对氨苄青霉素、羧苄青霉素和相关抗生素产生耐药性"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
AmpR promoter complement(13692..13796)
/gene="bla"
