| 质粒名称: | pAAVLINKv1-U6-sgRNA-EF1alpha-Cre-3'-SpCas9 |
|---|---|
| 出品公司: | Wekwikgene |
| 目录编号: | 0001911 |
| 产品用途: | 3' AAVLINK plasmid for SpCas9 expression |
| 存储实验室: | Zhonghua Lu Lab |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV-MCS2 |
| 表达系统: | 腺病毒 |
| 启动子: | EF1alpha |
| 拷贝数: | 高拷贝 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5alpha |
|
插入基因信息 |
|
| 插入基因名称: |
SpCas9
|
| 物种来源: | 未知 |
| 密码子优化: | 未优化 |
|
文献引用方法 |
|
| 参考文献部分: |
AAVLINK: A Potent DNA-Recombination Method for Large Cargo Delivery in Gene Therapy
Jianbang Lin, Yunping Lin, Sijia Wen, Wenhua Cao, Jianqing Zhang, Yujing Zhang, Zexuan Hong, Wenhui Liao, Yunyi Lin, Hanhe Liu, Qiwei Liu, Qi Li, Baiming Chen, Mengqi Li, Ziwei Luo, Luyu Yang, Yi Yang, Stephanie H. Zheng, Youcui Wang, Hongyuan Chu, Yu Hu, Yifan Qin, Brooke X. Luo, Shiyu Tian, Yefei Chen, Ting Yan, Lixin Yang, Hong Wang, Taian Liu, Yuwu Jiang, and Zhonghua Lu.
Under Review
|
pAAVLINKv1-U6-sgRNA-EF1alpha-Cre-3'-SpCas9
价格：元
规格： 2ug质粒（仅用于转化提取） 转染级去内毒素液体即用质粒100μg 转染级去内毒素液体即用质粒500μg 转染级去内毒素液体即用质粒1mg 更大规格-质粒构建-质粒定制-请联系我们
联系方式：I47-825O-882O
买家导航
pAAVLINKv1-U6-sgRNA-EF1alpha-Cre-3'-SpCas9载体质粒基本信息
pAAVLINKv1-U6-sgRNA-EF1alpha-Cre-3'-SpCas9质粒图谱载体图谱和pAAVLINKv1-U6-sgRNA-EF1alpha-Cre-3'-SpCas9载体序列质粒序列多克隆位点信息
pAAVLINKv1-U6-sgRNA-EF1alpha-Cre-3'-SpCas9质粒载体简介
pAAVLINKv1-U6-sgRNA-EF1alpha-Cre-3'-SpCas9质粒序列载体序列
LOCUS pAAVLINKv1-U6-sgRNA-EF1alpha-Cre-3'-SpCas9 7063 bp DNA circular SYN 10-AUG-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7063)
AUTHORS Stavrini Papadaki
TITLE Direct Submission
JOURNAL Exported Jun 15, 2026 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7063
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..130
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
/note="color: #ffe4c4"
promoter 145..393
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
promoter 145..385
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 415..490
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 510..1052
/label=EF1-alpha
/note="Chimeric promoter of ef1a and htlv 5' UTR"
promoter 539..750
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 763..1031
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
misc_feature 857..862
/label=SDS
CDS 1059..2090
/codon_start=1
/product="site-specific recombinase"
/label=Cre
/note="Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP sequences."
/translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
DGD"
intron 2091..2131
/label=Splicing Donor
protein_bind 2136..2169
/label=lox2272-JTZ17
/bound_moiety="Cre recombinase"
/note="Right element (RE) mutant of loxP (Araki et al.,
2010). Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
intron 2170..2261
/label=Splicing Acceptor
/note="Splicing acceptor"
CDS 2262..3800
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="VLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNL
TKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLK
SKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDV
RKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKG
RDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDS
PTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLI
IKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLF
TLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
CDS 2403..3800
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="AERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDEND
KLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLES
EFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIET
NGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKK
DWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLE
AKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHY
EKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKP
IREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI
DLSQLGGD"
CDS 3150..3800
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="GKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQK
QLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFT
LTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
CDS 3801..3848
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
terminator 3852..4283
/label=W3SL
/note="Short WPRE and SV40 polyA"
misc_feature 3996..4001
/label=SDS
repeat_region 4326..4466
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
misc_feature 4326..4455
/label=AAV2 ITR
repeat_region 4327..4467
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
rep_origin 4541..4996
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5278..5382
/gene="bla"
/label=AmpR promoter
CDS 5383..6243
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 6414..7002
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgtgagggc ctatttccca tgattccttc atatttgcat
181 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
241 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
301 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
361 gctttatata tcttgtggaa aggacgaaac accgactcca gtctttctag aagagtttta
421 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
481 gagtcggtgc tttttttcta gaggtaccgg atctgcgatc gctccggtgc ccgtcagtgg
541 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg
601 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
661 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
721 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
781 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
841 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
901 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
961 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
1021 cgttacagat ccaagctgtg accggcgcct accgtacgat gtccaattta ctgaccgtac
1081 accaaaattt gcctgcatta ccggtcgatg caacgagtga tgaggttcgc aagaacctga
1141 tggacatgtt cagggatcgc caggcgtttt ctgagcatac ctggaaaatg cttctgtccg
1201 tttgccggtc gtgggcggca tggtgcaagt tgaataaccg gaaatggttt cctgcagaac
1261 ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg cggtctggca gtaaaaacta
1321 tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg gtccgggctg ccacgaccaa
1381 gtgacagcaa tgctgtttca ctggttatgc ggcggatccg aaaagaaaac gttgatgccg
1441 gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga tttcgaccag gttcgttcac
1501 tcatggaaaa tagcgatcgc tgccaggata tacgtaatct ggcatttctg gggattgctt
1561 ataacaccct gttacgtata gccgaaattg ccaggatcag ggttaaagat atctcacgta
1621 ctgacggtgg gagaatgtta atccatattg gcagaacgaa aacgctggtt agcaccgcag
1681 gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt cgagcgatgg atttccgtct
1741 ctggtgtagc tgatgatccg aataactacc tgttttgccg ggtcagaaaa aatggtgttg
1801 ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct ggaagggatt tttgaagcaa
1861 ctcatcgatt gatttacggc gctaaggatg actctggtca gagatacctg gcctggtctg
1921 gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg cgctggagtt tcaataccgg
1981 agatcatgca agctggtggc tggaccaatg taaatattgt aatgaactat atccgtaacc
2041 tggatagtga aacaggggca atggtgcgcc tgctggaaga tggcgattag gtaagtatca
2101 aggttacaag acaggtttaa ggagaccaat agcttataac ttcgtatagg atactttata
2161 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
2221 tattggtctt actgacatcc actttgcctt tctctccaca ggtgctgacc agaagcgaca
2281 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
2341 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
2401 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
2461 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
2521 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
2581 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
2641 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
2701 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
2761 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
2821 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
2881 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
2941 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
3001 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
3061 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
3121 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
3181 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
3241 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
3301 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
3361 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
3421 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
3481 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
3541 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
3601 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
3661 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
3721 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
3781 tgtctcagct gggaggcgac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
3841 agaaaaagta actcgagata atcaacctct ggattacaaa atttgtgaaa gattgactgg
3901 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgca
3961 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt
4021 tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
4081 gttgggcact gacaattccg tggtgtttat ttgtgaaatt tgtgatgcta ttgctttatt
4141 tgtaaccatc tagctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
4201 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
4261 gggagatgtg ggaggttttt taaagatctg gagaccggtg tggcacgtgc ggaccgagcg
4321 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
4381 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
4441 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
4501 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
4561 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
4621 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
4681 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
4741 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
4801 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
4861 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
4921 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
4981 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
5041 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
5101 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
5161 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
5221 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
5281 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
5341 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
5401 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
5461 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
5521 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
5581 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
5641 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
5701 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
5761 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
5821 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
5881 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
5941 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
6001 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
6061 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
6121 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
6181 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
6241 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
6301 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
6361 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
6421 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
6481 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
6541 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
6601 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
6661 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
6721 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
6781 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
6841 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
6901 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
6961 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
7021 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
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