| 质粒名称: | pU6-pegRNA-GG-acceptor |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 132777 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pU6 |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 哺乳细胞表达 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Exchangeable cassette,mCherry |
| 物种来源: | Synthetic |
| 插入大小: | 821 bp |
| 基因突变: | See manuscript |
| 启动子 | U6 |
| 克隆方法: | Ligation Independent克隆 |
| 5' 测序引物 | GACTATCATATGCTTACCGT |
|
文献引用方法 |
|
| 材料和方法部分: | pU6-pegRNA-GG-acceptor was a gift from David Liu (Addgene plasmid # 132777 ; http://n2t.net/addgene:132777 ; RRID:Addgene_132777) |
| 参考文献部分: | Search-and-replace genome editing without double-strand breaks or donor DNA. Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, Chen PJ, Wilson C, Newby GA, Raguram A, Liu DR. Nature. 2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 10.1038/s41586-019-1711-4. 10.1038/s41586-019-1711-4 PubMed 31634902 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pU6-pegRNA-GG-acceptor 3004 bp ds-DNA circular SYN 18-OCT-2019
DEFINITION Prime editing in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pU6-pegRNA-GG-acceptor
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3004)
TITLE Liu Lab plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 3004)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 18, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3004
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
terminator 259..290
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
CDS complement(307..984)
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
RBS 991..1002
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
protein_bind 1020..1038
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1045..1063
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
rep_origin complement(1174..1762)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1254..1273)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1933..2793)
/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 2556..2575
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2794..2898)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccc gagaccgtag tcaaaagcct ccggtcggag gcttttgact tggctgagga
301 gtgccattaa gcaccggtgg agtgacgacc ttcagcacgt tcgtactgtt caacgatggt
361 gtagtcttcg ttgtgggagg tgatgtccag tttgatgtcg gttttgtaag cacccggcag
421 ctgaaccggt tttttagcca tgtaggtggt tttaacttca gcgtcgtagt gaccaccgtc
481 tttcagtttc agacgcattt tgatttcacc tttcagagca ccgtcttccg ggtacatacg
541 ttcggtggaa gcttcccaac ccatggtttt tttctgcata accggaccgt cggacgggaa
601 gttggtacca cgcagtttaa ctttgtagat gaactcaccg tcttgcaggg aggagtcctg
661 ggtaacggta acaacaccac cgtcttcgaa gttcataaca cgttcccatt tgaaaccttc
721 cgggaaggac agtttcaggt agtccgggat gtcagccggg tgtttaacgt aagctttgga
781 accgtactgg aactgcgggg acaggatgtc ccaagcgaac ggcagcggac cacctttggt
841 aactttcagt ttagcggtct gggtaccttc gtacggacga ccttcacctt caccttcgat
901 ttcgaactcg tgaccgttaa cggaaccttc catacgaact ttgaaacgca tgaactcttt
961 gataacgtct tcgctactcg ccatagatcc tttctcctct ttcagatccg tgctcagtat
1021 ctctatcact gatagggatg tcaatctcta tcactgatag ggaggtctcg tttttttaag
1081 cttgggccgc tcgaggtacc tctctacata tgacatgtga gcaaaaggcc agcaaaaggc
1141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
1201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
1261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
1321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
1381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
1441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
1501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
1561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
1621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
1681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
1741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
1801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
1861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
1921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
1981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
2041 accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt
2101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
2161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
2221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
2281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
2341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
2401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
2461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
2521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
2581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
2641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
2701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
2761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
2821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
2881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg ctagctgtac
2941 aaaaaagcag gctttaaagg aaccaattca gtcgactgga tccggtacca aggtcgggca
3001 ggaa
//
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