| 质粒名称: | pPEgRNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 172716 |
| 存储实验室: | Tilmann Weber |
|
载体骨架基本信息 |
|
| 载体名称: | pgRNA-bacteria |
| 载体出品公司: | Clontech |
| 空载体大小: | 2488 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: |
gRNA
gRNA/shRNA sequencescaffold: gttttagagctagaaatagcaagttaaaataaggctagtccgttatcaacttgaaaaagtggcaccgagtcggtgc
|
| 物种来源: | Synthetic |
| 插入大小: | 76bp |
| 克隆方法: | Gibson Cloning |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pPEgRNA was a gift from Tilmann Weber (Addgene plasmid # 172716 ; http://n2t.net/addgene:172716 ; RRID:Addgene_172716) |
| 参考文献部分: | A versatile genetic engineering toolkit for E. coli based on CRISPR-prime editing. Tong Y, Jorgensen TS, Whitford CM, Weber T, Lee SY. Nat Commun. 2021 Sep 1;12(1):5206. doi: 10.1038/s41467-021-25541-3. 10.1038/s41467-021-25541-3 PubMed 34471126 |
买家导航
pPEgRNA载体质粒基本信息
pPEgRNA质粒图谱载体图谱和pPEgRNA载体序列质粒序列多克隆位点信息
pPEgRNA质粒载体简介
pPEgRNA质粒序列载体序列
LOCUS pPEgRNA 2564 bp DNA circular SYN 25-AUG-2024
DEFINITION Delivery of guide RNA for prime editing.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2564)
AUTHORS Tong Y, Jorgensen TS, Whitford CM, Weber T, Lee SY
TITLE A versatile genetic engineering toolkit for E. coli based on
CRISPR-prime editing.
JOURNAL Nat Commun. 2021 Sep 1;12(1):5206. doi: 10.1038/s41467-021-25541-3.
PUBMED 34471126
REFERENCE 2 (bases 1 to 2564)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun."; date: "2021-09-1"; volume: "12(1):5206. doi"; pages: "
10.1038/s41467-021-25541-3"
FEATURES Location/Qualifiers
source 1..2564
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 37..71
/label=J23119(SpeI) promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119) modified to end with an SpeI site"
misc_RNA 72..147
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 168..197
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 213..230
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(286..303)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 456..502
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind complement(555..572)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(726..1314)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(806..825)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1485..2345)
/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 2108..2127
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2346..2450)
/gene="bla"
/label=AmpR promoter
primer_bind 2518..2536
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator complement(join(2540..2564,1..19))
/label=bacterial terminator
/note="putative bacterial transcription terminator"
ORIGIN
1 tttcgctaag gatgatttct ggaattctaa agatctttga cagctagctc agtcctaggt
61 ataatactag tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
121 cttgaaaaag tggcaccgag tcggtgcttt ttttgaagct tgggcccgaa caaaaactca
181 tctcagaaga ggatctgaat agcgccgtcg accatcatca tcatcatcat tgagtttaaa
241 cggtctccag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
301 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
361 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
421 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
481 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa ctggatcctt actcgagtct
541 agactgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
601 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
661 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
721 gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
781 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
841 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
901 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
961 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
1021 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
1081 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
1141 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
1201 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
1261 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
1321 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
1381 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
1441 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
1501 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
1561 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
1621 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
1681 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
1741 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
1801 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
1861 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
1921 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
1981 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
2041 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
2101 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
2161 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
2221 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
2281 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
2341 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
2401 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
2461 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
2521 aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagc
//
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