| 质粒名称: | pAC1372-pX-sgRNA-15xPBSa |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 71889 |
| 存储实验室: | Albert Cheng |
|
载体骨架基本信息 |
|
| 载体名称: | pX330; pUC ori vector |
| 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 |
|
插入基因信息 |
|
| 5’测序引物 | gagggcctatttcccatgattcc |
| 3’测序引物 | gccatttgtctgcagaattggc |
|
文献引用方法 |
|
| 材料和方法部分: | pAC1372-pX-sgRNA-15xPBSa was a gift from Albert Cheng (Addgene plasmid # 71889 ; http://n2t.net/addgene:71889 ; RRID:Addgene_71889) |
| 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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载体基本信息
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载体简介
载体序列
LOCUS pAC1372-pX-sgRNA-15xPBSa 3381 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Cloning vector for expression of sgRNA-15xPBSa.
ACCESSION .
VERSION .
KEYWORDS pAC1372-pX-sgRNA-15xPBSa
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3381)
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 3381)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3381
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 9..249
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
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 644..784
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 644..773
/label=AAV2 ITR
rep_origin 859..1314
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(946..965)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1156..1177
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1331..1350)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1450..1472
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1510..1528)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1596..1700
/gene="bla"
/label=AmpR promoter
CDS 1701..2561
/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(1919..1938)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2732..3320
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3221..3240
/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 aaacaccggg tcttcgagaa gacctgttta agagctatgc tggaaacagc
301 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
361 ccaattgggt ctccagattg tatgtagcct gtatgtagcc tgtatgtagc ctgtatgtag
421 cctgtatgta agattgtatg tagctgtatg tagcctgtat gtagcctgta tgtagcctgt
481 atgtaagatt gtatgtagcc tgtatgtagc ctgtatgtag cctgtatgta gcctgtatgt
541 aagatctttt tttgttttag agctagaaat agcaagttaa aataaggcta gtccgtagcg
601 cgtgcgccaa ttctgcagac aaatggcggc gcgccgcggc cgcaggaacc cctagtgatg
661 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
721 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
781 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
841 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
901 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
961 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
1021 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
1081 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
1141 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
1201 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
1261 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
1321 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
1381 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
1441 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
1501 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
1561 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
1621 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
1681 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
1741 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
1801 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
1861 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
1921 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
1981 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
2041 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
2101 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
2161 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
2221 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
2281 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
2341 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
2401 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
2461 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
2521 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
2581 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
2641 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
2701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
2761 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
2821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
2881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
2941 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
3001 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
3061 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
3121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
3181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
3241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
3301 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
3361 gctggccttt tgctcacatg t
//
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