| 质粒名称: | pDY0663 ACTB atgRNA with v2 scaffold |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 179108 |
| 存储实验室: | Omar Abudayyeh, Jonathan Gootenberg |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 2377bp |
| 载体类型: | CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | ACTB N-term PBS 13 RT 29 AttB 46 atgRNA with v2 scaffold |
|
文献引用方法 |
|
| 材料和方法部分: | pDY0663 ACTB atgRNA with v2 scaffold was a gift from Omar Abudayyeh & Jonathan Gootenberg (Addgene plasmid # 179108 ; http://n2t.net/addgene:179108 ; RRID:Addgene_179108) |
| 参考文献部分: | 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载体质粒基本信息
pDY0663 ACTB atgRNA with v2 scaffold质粒图谱载体图谱和pDY0663 ACTB atgRNA with v2 scaffold载体序列质粒序列多克隆位点信息
pDY0663 ACTB atgRNA with v2 scaffold质粒载体简介
pDY0663 ACTB atgRNA with v2 scaffold质粒序列载体序列
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
https://www.biofeng.com
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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