质粒名称: | pAC1454_pX-sgCentromere-25xPBSc |
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出品公司: | Addgene |
目录编号: | 73696 |
存储实验室: | Albert Cheng |
载体骨架基本信息 |
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载体名称: | pX330; pUC ori vector |
载体出品公司: | Clontech |
完整质粒大小: | 3441bp |
载体类型: | 哺乳细胞表达, CRISPR |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: |
sgCentromere-20xPBSc
gRNA/shRNA sequenceGTTGAGGCCTTCGTTGGAAAC
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启动子: | U6 |
克隆方法: | WZ |
5’测序引物 | gagggcctatttcccatgattcc |
3’测序引物 | gccatttgtctgcagaattggc |
文献引用方法 |
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材料和方法部分: | pAC1454_pX-sgCentromere-25xPBSc was a gift from Albert Cheng (Addgene plasmid # 73696) |
参考文献部分: | Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, Taghbalout A, Wang H. Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15. 10.1038/cr.2016.3 PubMed 26768771 |
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pAC1454_pX-sgCentromere-25xPBSc载体质粒基本信息
pAC1454_pX-sgCentromere-25xPBSc质粒图谱载体图谱和pAC1454_pX-sgCentromere-25xPBSc载体序列质粒序列多克隆位点信息
pAC1454_pX-sgCentromere-25xPBSc质粒载体简介
pAC1454_pX-sgCentromere-25xPBSc质粒序列载体序列
LOCUS pAC1454_pX-sgCentromere-25xPBSc 3441 bp DNA circular SYN 26-AUG-2024 DEFINITION Vector expression sgCentromere-20xPBSc. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3441) AUTHORS Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, Taghbalout A, Wang H TITLE Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. JOURNAL Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15. PUBMED 26768771 REFERENCE 2 (bases 1 to 3441) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.biofeng.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/cr.2016.3"; journalName: "Cell Res"; date: "2016-02"; volume: "26"; issue: "2"; pages: "254-7" FEATURES Location/Qualifiers source 1..3441 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 9..257 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)" primer_bind 9..29 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 180..199 /label=LKO.1 5' /note="Human U6 promoter, forward primer" repeat_region 704..844 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 704..833 /label=AAV2 ITR rep_origin 919..1374 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1006..1025) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1216..1237 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1391..1410) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1510..1532 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1570..1588) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1656..1760 /gene="bla" /label=AmpR promoter CDS 1761..2621 /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(1979..1998) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2792..3380 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3281..3300 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ggcgcgccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg 61 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt 121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg 181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt 241 ggaaaggacg aaacaccgtt gaggccttcg ttggaaacgt ttaagagcta tgctggaaac 301 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg 361 gtgccaattg ggtctccaga tttgatgtag ccttgatgta gccttgatgt agccttgatg 421 tagccttgat gtaagatttg atgtagcctt gatgtagcct tgatgtagcc ttgatgtagc 481 cttgatgtaa gatttgatgt agccttgatg tagccttgat gtagccttga tgtagccttg 541 atgtaagatt tgatgtagcc ttgatgtagc cttgatgtag ccttgatgta gccttgatgt 601 aagatctttt tttgttttag agctagaaat agcaagttaa aataaggcta gtccgtagcg 661 cgtgcgccaa ttctgcagac aaatggcggc gcgccgcggc cgcaggaacc cctagtgatg 721 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc 781 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg 841 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 901 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 961 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 1021 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 1081 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg 1141 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 1201 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 1261 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 1321 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt 1381 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa 1441 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 1501 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 1561 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 1621 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 1681 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 1741 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 1801 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 1861 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 1921 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 1981 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 2041 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 2101 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 2161 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 2221 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 2281 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 2341 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 2401 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 2461 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct 2521 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 2581 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 2641 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 2701 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 2761 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 2821 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 2881 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc 2941 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 3001 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 3061 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 3121 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 3181 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 3241 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 3301 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 3361 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 3421 gctggccttt tgctcacatg t //
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