| 质粒名称: | AAV-U6gRNA1-U6gRNA2-TnT-Cre |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 87682 |
| 存储实验室: | William Pu |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2900 bp |
| 完整质粒大小: | 5969 bp |
| 载体类型: | AAV表达 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | gRNA1 |
| 插入大小: | 102 bp |
| 启动子 | U6 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | KpnI (未破坏) |
| 3' 克隆位点: | MluI(未破坏) |
| 5' 测序引物 | CGGCCGCACGCGCCGGTACC |
| 3' 测序引物 | CAGAAGAGCTCGCTCTTCCG |
|
文献引用方法 |
|
| 材料和方法部分: | AAV-U6gRNA1-U6gRNA2-TnT-Cre was a gift from William Pu (Addgene plasmid # 87682 ; http://n2t.net/addgene:87682 ; RRID:Addgene_87682) |
| 参考文献部分: | Analysis of Cardiac Myocyte Maturation Using CASAAV, A Platform for Rapid Dissection of Cardiac Myocyte Gene Function In Vivo. Guo Y, VanDusen NJ, Zhang L, Gu W, Sethi I, Guatimosim S, Ma Q, Jardin BD, Ai Y, Zhang D, Chen B, Guo A, Yuan GC, Song LS, Pu WT. Circ Res. 2017 Mar 29. pii: CIRCRESAHA.116.310283. doi: 10.1161/CIRCRESAHA.116.310283. 10.1161/CIRCRESAHA.116.310283 PubMed 28356340 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS AAV-U6gRNA1-U6gRNA2-TnT-Cre 5958 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION AAV vector for U6 driven expression of two gRNAs, and cardiomyocyte
specific expression of Cre recombinase..
ACCESSION .
VERSION .
KEYWORDS AAV-U6gRNA1-U6gRNA2-TnT-Cre
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5958)
AUTHORS Guo Y, VanDusen NJ, Zhang L, Gu W, Sethi I, Guatimosim S, Ma Q,
Jardin BD, Ai Y, Zhang D, Chen B, Guo A, Yuan GC, Song LS, Pu WT
TITLE Analysis of Cardiac Myocyte Maturation Using CASAAV, A Platform for
Rapid Dissection of Cardiac Myocyte Gene Function In Vivo.
JOURNAL Circ Res. 2017 Mar 29. pii: CIRCRESAHA.116.310283. doi:
10.1161/CIRCRESAHA.116.310283.
PUBMED 28356340
REFERENCE 2 (bases 1 to 5958)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5958
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 278..353
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 366..606
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 633..708
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
intron 1260..1392
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 1375..1394
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 1437..1455
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1467..1487
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 2554..3030
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2753..2772)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3070..3210
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3070..3199
/label=AAV2 ITR
rep_origin 3285..3740
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3372..3391)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3582..3603
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3757..3776)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3876..3898
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3936..3954)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4022..4126
/gene="bla"
/label=AmpR promoter
CDS 4127..4987
/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"
primer_bind complement(4345..4364)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5158..5746
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5647..5666
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
repeat_region 5808..5937
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg actgcaggtg gaattccacc tgctcaggtt ttagagctag aaatagcaag
301 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttta
361 cgcgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
421 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
481 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
541 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
601 aaggacgaaa caccggaaga gcgagctctt ctgttttaga gctagaaata gcaagttaaa
661 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttctagt
721 gggataaaag cagtctgggc tttcacatga cagcatctgg ggctgcggca gagggtcggg
781 tccgaagcgc tgccttatca gcgtccccag ccctgggagg tgacagctgg ctggcttgtg
841 tcagcccctc gggcactcac gtatctccgt ccgacgggtt taaaatagca aaactctgag
901 gccacacaat agcttgggct tatatgggct cctgtggggg aagggggagc acggaggggg
961 ccggggccgc tgctgccaaa atagcagctc acaagtgttg cattcctctc tgggcgccgg
1021 gcacattcct gctggctctg cccgccccgg ggtgggcgcc ggggggacct taaagcctct
1081 gccccccaag gagcccttcc cagacagccg ccggcaccca ccgctccgtg ggacgatccc
1141 cgaagctcta gagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
1201 cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg
1261 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
1321 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
1381 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat
1441 acgactcact ataggctagc atggtgccca agaagaagag gaaagtctcc aacctgctga
1501 ctgtgcacca aaacctgcct gccctccctg tggatgccac ctctgatgaa gtcaggaaga
1561 acctgatgga catgttcagg gacaggcagg ccttctctga acacacctgg aagatgctcc
1621 tgtctgtgtg cagatcctgg gctgcctggt gcaagctgaa caacaggaaa tggttccctg
1681 ctgaacctga ggatgtgagg gactacctcc tgtacctgca agccagaggc ctggctgtga
1741 agaccatcca acagcacctg ggccagctca acatgctgca caggagatct ggcctgcctc
1801 gcccttctga ctccaatgct gtgtccctgg tgatgaggag aatcagaaag gagaatgtgg
1861 atgctgggga gagagccaag caggccctgg cctttgaacg cactgacttt gaccaagtca
1921 gatccctgat ggagaactct gacagatgcc aggacatcag gaacctggcc ttcctgggca
1981 ttgcctacaa caccctgctg cgcattgccg aaattgccag aatcagagtg aaggacatct
2041 cccgcaccga tggtgggaga atgctgatcc acattggcag gaccaagacc ctggtgtcca
2101 cagctggtgt ggagaaggcc ctgtccctgg gggttaccaa gctggtggag agatggatct
2161 ctgtgtctgg tgtggctgat gaccccaaca actacctgtt ctgccgggtc agaaagaatg
2221 gtgtggctgc cccttctgcc acctcccaac tgtccacccg ggccctggaa gggatctttg
2281 aggccaccca ccgcctgatc tatggtgcca aggatgactc tgggcagaga tacctggcct
2341 ggtctggcca ctctgccaga gtgggtgctg ccagggacat ggccagggct ggtgtgtcca
2401 tccctgaaat catgcaggct ggtggctgga ccaatgtgaa cattgtgatg aactacatca
2461 gaaacctgga ctctgagact ggggccatgg tgaggctgct cgaggatggg gactgaaagc
2521 ttatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
2581 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
2641 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
2701 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
2761 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
2821 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
2881 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
2941 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
3001 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
3061 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
3121 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
3181 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
3241 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
3301 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
3361 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
3421 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
3481 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
3541 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
3601 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
3661 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
3721 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
3781 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
3841 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
3901 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
3961 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
4021 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
4081 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
4141 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
4201 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
4261 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
4321 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
4381 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
4441 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
4501 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
4561 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
4621 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
4681 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
4741 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
4801 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
4861 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
4921 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
4981 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
5041 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
5101 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
5161 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
5221 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
5281 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
5341 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
5401 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
5461 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
5521 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
5581 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
5641 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
5701 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
5761 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
5821 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
5881 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
5941 gccgcacgcg ccggtacc
//
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