| 質粒名稱: | pDY0663 ACTB atgRNA with v2 scaffold |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 179108 |
| 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: | 2377bp |
| 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: | 未知 |
|
插入基因信息 |
|
| 插入基因名稱: | ACTB N-term PBS 13 RT 29 AttB 46 atgRNA with v2 scaffold |
|
文獻引用方法 |
|
| 材料和方法部分: | pDY0663 ACTB atgRNA with v2 scaffold was a gift from Omar Abudayyeh & Jonathan Gootenberg (Addgene plasmid # 179108 ; //n2t.net/addgene:179108 ; RRID:Addgene_179108) |
| 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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pDY0663 ACTB atgRNA with v2 scaffold載體質粒基本信息
pDY0663 ACTB atgRNA with v2 scaffold質粒圖譜載體圖譜和pDY0663 ACTB atgRNA with v2 scaffold載體序列質粒序列多克隆位點信息
pDY0663 ACTB atgRNA with v2 scaffold質粒載體簡介
pDY0663 ACTB atgRNA with v2 scaffold質粒序列載體序列
LOCUS pDY0663 ACTB atgRNA with v2 scaffold 2377 bp DNA circular SYN 14-APR-2025
DEFINITION ACTB N-term PBS 13 RT 29 Bxb1 AttB 46 atgRNA with v2 scaffold.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2377)
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 2377)
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: pDY0663 ACTB atgRNA with v2 scaffold
FEATURES Location/Qualifiers
source 1..2377
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(38..626)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(118..137)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(797..1588,1589..1657))
/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 1420..1439
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1658..1762)
/gene="bla"
/label=AmpR promoter
promoter 1869..2117
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 1869..1889
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2040..2059
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
protein_bind 2257..2294
/label=Bxb1 attB
/bound_moiety="Mycobacteriophage Bxb1 integrase"
/note="Minimal attB site for the Mycopbacteriophage Bxb1
integrase (Voutev and Mann, 2017)"
ORIGIN
1 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
61 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
421 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
481 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
541 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
721 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
901 gcttaccatc tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag
961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
1021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
1081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
1141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
1201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
1261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
1321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
1381 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
1441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
1501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
1561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
1621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
1681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
1741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgctagct
1801 gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac tggatccggt accaaggtcg
1861 ggcaggaaga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
1921 ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt
1981 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
2041 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
2101 ggaaaggacg aaacaccgct attctcgcag ctcaccagtt tgagagctat gctggaaaca
2161 gcatagcaag ttcaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
2221 tgcgacgagc gcggcgatat catcatccat ggccggatga tcctgacgac ggagaccgcc
2281 gtcgtcgaca agccggcctg agctgcgaga atttttttaa gcttgggccg ctcgaggtac
2341 ctctctacat atgacatgtg agcaaaaggc cagcaaa
//
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