载体基本信息 | |||
载体名称 | AAV-U6gRNA1-U6gRNA2-TnT-Cre | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 5958 bp | 载体类型 | AAV |
启动子 | cTnT | 克隆方法 | Restriction Enzyme |
5'引物 | ACGACTCACTATAGGCTAGC | 3'引物 | AGGGACTTCGGGCACAATCG |
AAV-U6gRNA1-U6gRNA2-TnT-Cre 载体载体图谱
AAV-U6gRNA1-U6gRNA2-TnT-Cre 载体载体序列
LOCUS Exported 5958 bp ds-DNA circular SYN 13-MAY-2021 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 COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Circ Res. 2017 Mar 29. pii: CIRCRESAHA.116.310283. doi: 10.1161/CIRCRESAHA.116.310283." 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"
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