| 質粒名稱: | pDY0209 ACTB Nicking guide +48 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 179109 |
| 存儲實驗室: | Omar Abudayyeh, Jonathan Gootenberg |
|
載體骨架基本信息 |
|
| 載體名稱: | pCMV |
| 載體出品公司: | Clontech |
| 完整質粒大小: | 2279bp |
| 載體類型: | CRISPR |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | ACTB N-term Nicking guide 1 +48 guide |
|
文獻引用方法 |
|
| 材料和方法部分: | pDY0209 ACTB Nicking guide +48 was a gift from Omar Abudayyeh & Jonathan Gootenberg (Addgene plasmid # 179109 ; //n2t.net/addgene:179109 ; RRID:Addgene_179109) |
| 參考文獻部分: | Drag-and-drop genome insertion of large sequences without double-strand DNA cleavage using CRISPR-directed integrases. Yarnall MTN, Ioannidi EI, Schmitt-Ulms C, Krajeski RN, Lim J, Villiger L, Zhou W, Jiang K, Garushyants SK, Roberts N, Zhang L, Vakulskas CA, Walker JA 2nd, Kadina AP, Zepeda AE, Holden K, Ma H, Xie J, Gao G, Foquet L, Bial G, Donnelly SK, Miyata Y, Radiloff DR, Henderson JM, Ujita A, Abudayyeh OO, Gootenberg JS. Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4. 10.1038/s41587-022-01527-4 PubMed 36424489 |
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pDY0209 ACTB Nicking guide +48載體質粒基本信息
pDY0209 ACTB Nicking guide +48質粒圖譜載體圖譜和pDY0209 ACTB Nicking guide +48載體序列質粒序列多克隆位點信息
pDY0209 ACTB Nicking guide +48質粒載體簡介
pDY0209 ACTB Nicking guide +48質粒序列載體序列
LOCUS pDY0209 ACTB Nicking guide +48 2279 bp DNA circular SYN 14-APR-2025
DEFINITION ACTB N-term Nicking guide 1 +48 guide.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2279)
AUTHORS Yarnall MTN, Ioannidi EI, Schmitt-Ulms C, Krajeski RN, Lim J,
Villiger L, Zhou W, Jiang K, Garushyants SK, Roberts N, Zhang L,
Vakulskas CA, Walker JA 2nd, Kadina AP, Zepeda AE, Holden K, Ma H,
Xie J, Gao G, Foquet L, Bial G, Donnelly SK, Miyata Y, Radiloff DR,
Henderson JM, Ujita A, Abudayyeh OO, Gootenberg JS
TITLE Drag-and-drop genome insertion of large sequences without
double-strand DNA cleavage using CRISPR-directed integrases.
JOURNAL Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4.
PUBMED 36424489
REFERENCE 2 (bases 1 to 2279)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4."
COMMENT Sequence Label: pDY0209 ACTB Nicking guide +48
FEATURES Location/Qualifiers
source 1..2279
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 393..412
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(631..735)
/gene="bla"
/label=AmpR promoter
promoter 842..1090
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 842..862
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 1013..1032
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 1111..1186
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
rep_origin complement(1290..1878)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1370..1389)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(2049..2279,1..561,562..630))
/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"
ORIGIN
1 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
61 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
121 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
181 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
241 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
301 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
361 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
421 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
481 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
541 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
601 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
661 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
721 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgcta gctgtacaaa
781 aaagcaggct ttaaaggaac caattcagtc gactggatcc ggtaccaagg tcgggcagga
841 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
901 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
961 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
1021 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
1081 acgaaacacc gaagccggcc ttgcacatgc gttttagagc tagaaatagc aagttaaaat
1141 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttaagcttg
1201 ggccgctcga ggtacctctc tacatatgac atgtgagcaa aaggccagca aaaggccagg
1261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
1321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
1381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
1441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
1501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
1561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
1621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
1681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
1741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
1801 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
1861 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
1921 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
1981 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
2041 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
2101 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
2161 tctggcccca gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca
2221 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgc
//