| 质粒名称: | pC0052-REPAIR non-targeting guide clone into pC0043 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 103868 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pC0043-PspCas13b crRNA backbone |
| 空载体大小: | 2962 bp |
| 完整质粒大小: | 2970 bp |
| 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: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | non-targeting guide RNA |
| 5’测序引物 | GTAATGCCTGGCTTGTCGACGCATAGTCTG |
| 3’测序引物 | GTAATGCCTGGCTTGTCGACGCATAGTCTG |
|
文献引用方法 |
|
| 材料和方法部分: | pC0052-REPAIR non-targeting guide clone into pC0043 was a gift from Feng Zhang (Addgene plasmid # 103868 ; http://n2t.net/addgene:103868 ; RRID:Addgene_103868) |
| 参考文献部分: | RNA editing with CRISPR-Cas13. Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung J, Zhang F. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. 10.1126/science.aaq0180 PubMed 29070703 |
pC0052-REPAIR non-targeting guide clone into pC0043
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载体基本信息
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载体简介
载体序列
LOCUS pC0052-REPAIR non-targeting guide clone into pC0043 2970 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Non-targeting guide RNA for use with PspCas13b. Useful as a control
for RNA editing and RNA knockdown experiments..
ACCESSION .
VERSION .
KEYWORDS pC0052-REPAIR non-targeting guide clone into pC0043
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2970)
AUTHORS Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung
J, Zhang F
TITLE RNA editing with CRISPR-Cas13.
JOURNAL Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
PUBMED 29070703
REFERENCE 2 (bases 1 to 2970)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..2970
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 112..129
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 246..267
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 282..312
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 320..336
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 344..360
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 344..360
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 374..614
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 374..394
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 545..564
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(714..731)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(714..730)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(723..745)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(939..958)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1058..1080
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1118..1136)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1204..1308
/gene="bla"
/label=AmpR promoter
CDS 1309..2169
/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(1527..1546)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2340..2928
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2829..2848
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
61 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
121 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
181 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
241 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
301 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acgcaggaaa cagctatgac
361 catgattacg ccagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
421 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
481 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
541 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
601 cttgtggaaa ggacgaaaca ccggtaatgc ctggcttgtc gacgcatagt ctggttgtgg
661 aaggtccagt tttgaggggc tattacaact tttttggtac cgagctcgaa ttcactggcc
721 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
781 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
841 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
901 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
961 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
1021 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
1081 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
1141 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
1201 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
1261 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
1321 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
1381 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
1441 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
1501 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
1561 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
1621 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
1681 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
1741 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
1801 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
1861 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
1921 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
1981 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
2041 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
2101 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
2161 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
2221 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
2281 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
2341 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
2401 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
2461 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
2521 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
2581 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
2641 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
2701 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
2761 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
2821 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
2881 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
2941 gcggcctttt tacggttcct ggccttttgc
//
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