| 质粒名称: | pU6-tevopreq1-GG-acceptor |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 174038 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pBR322 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3041 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | hU6 |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pU6-tevopreq1-GG-acceptor was a gift from David Liu (Addgene plasmid # 174038 ; http://n2t.net/addgene:174038 ; RRID:Addgene_174038) |
| 参考文献部分: | Engineered pegRNAs improve prime editing efficiency. Nelson JW, Randolph PB, Shen SP, Everette KA, Chen PJ, Anzalone AV, An M, Newby GA, Chen JC, Hsu A, Liu DR. Nat Biotechnol. 2021 Oct 4. pii: 10.1038/s41587-021-01039-7. doi: 10.1038/s41587-021-01039-7. 10.1038/s41587-021-01039-7 PubMed 34608327 |
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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-tevopreq1-GG-acceptor 3041 bp ds-DNA circular SYN 17-AUG-2021
DEFINITION Prime editing in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pU6-tevopreq1-GG-acceptor
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3041)
TITLE Cloning vectors for prime editing guide RNAs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 3041)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 17, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3041
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 82..322
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 82..102
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 253..272
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
terminator 340..371
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
CDS complement(388..1065)
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
RBS 1072..1083
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
protein_bind 1101..1119
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1126..1144
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
rep_origin complement(1292..1880)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1372..1391)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2051..2911)
/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 2674..2693
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2912..3016)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
301 ttatatatct tgtggaaagg acgaaacacc cgagaccgta gtcaaaagcc tccggtcgga
361 ggcttttgac ttggctgagg agtgccatta agcaccggtg gagtgacgac cttcagcacg
421 ttcgtactgt tcaacgatgg tgtagtcttc gttgtgggag gtgatgtcca gtttgatgtc
481 ggttttgtaa gcacccggca gctgaaccgg ttttttagcc atgtaggtgg ttttaacttc
541 agcgtcgtag tgaccaccgt ctttcagttt cagacgcatt ttgatttcac ctttcagagc
601 accgtcttcc gggtacatac gttcggtgga agcttcccaa cccatggttt ttttctgcat
661 aaccggaccg tcggacggga agttggtacc acgcagttta actttgtaga tgaactcacc
721 gtcttgcagg gaggagtcct gggtaacggt aacaacacca ccgtcttcga agttcataac
781 acgttcccat ttgaaacctt ccgggaagga cagtttcagg tagtccggga tgtcagccgg
841 gtgtttaacg taagctttgg aaccgtactg gaactgcggg gacaggatgt cccaagcgaa
901 cggcagcgga ccacctttgg taactttcag tttagcggtc tgggtacctt cgtacggacg
961 accttcacct tcaccttcga tttcgaactc gtgaccgtta acggaacctt ccatacgaac
1021 tttgaaacgc atgaactctt tgataacgtc ttcgctactc gccatagatc ctttctcctc
1081 tttcagatcc gtgctcagta tctctatcac tgatagggat gtcaatctct atcactgata
1141 gggaggtctc gcgcggttct atctagttac gcgttaaacc aactagaatt tttttaagct
1201 tgggccgctc gaggtacctc tctacatatg acatgtgagc aaaaggccag caaaaggcca
1261 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
1321 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
1381 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
1441 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
1501 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
1561 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
1621 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
1681 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
1741 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
1801 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
1861 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
1921 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
1981 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
2041 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
2101 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
2161 catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat
2221 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
2281 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
2341 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
2401 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
2461 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
2521 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
2581 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
2641 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
2701 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
2761 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
2821 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
2881 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
2941 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
3001 aaataggggt tccgcgcaca tttccccgaa aagtgccacc t
//
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