| 质粒名称: | pEF1a-mMLH1dn |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 174825 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | Custom |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 6315bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | mouse MLH1dn (codon optimized) / mus musculus MLH1 del758-760 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2271bp |
| 基因突变: | eletion of residues 758-760 |
| 启动子: | EF1a |
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | EF1a-forward |
| 3’测序引物 | BGH-reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pEF1a-mMLH1dn was a gift from David Liu (Addgene plasmid # 174825 ; http://n2t.net/addgene:174825 ; RRID:Addgene_174825) |
| 参考文献部分: | Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, Liu DR. Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 10.1016/j.cell.2021.09.018. 10.1016/j.cell.2021.09.018 PubMed 34653350 |
买家导航
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 pEF1a-mMLH1dn 6315 bp ds-DNA circular SYN 27-SEP-2021
DEFINITION Mammalian expression of mouse MLH1dn (codon optimized).
ACCESSION .
VERSION .
KEYWORDS pEF1a-mMLH1dn
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6315)
TITLE Enhanced Prime Editing Systems Through Identification and
Manipulation of Cellular Determinants of Editing Outcomes
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6315)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6315
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 197..1375
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 428..1366
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1323..1343
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 1394..1403
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3679..3696
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3719..3736)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3725..3949
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(4020..4036)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4020..4036)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4033..4055)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4044..4060
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4068..4098)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4113..4134
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4251..4268)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4422..5010)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4502..4521)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5181..6041)
/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 5804..5823
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6042..6146)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6225..6244)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
61 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactgca
121 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
181 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
241 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
301 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
361 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
421 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
481 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
541 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
601 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
661 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
721 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
781 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
841 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
901 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
961 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
1021 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
1081 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
1141 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
1201 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
1261 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
1321 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgacgtac
1381 gggatcctct agagccacca tggcattcgt ggctggtgtc atccgccgtc ttgatgagac
1441 tgtcgtcaac agaattgccg caggagaagt catccagcgt ccagccaacg ccatcaagga
1501 aatgatcgaa aactgtctgg atgccaagag cacaaacata caagttgtgg tgaaggaggg
1561 tggcctcaaa ctcattcaga tccaagacaa cggcacggga attagaaaag aagacctgga
1621 cattgtgtgc gagcgcttca caacctctaa attgcagaca tttgaggacc tggcaagcat
1681 cagtacttat ggcttccgag gggaggctct ggcctccata agccatgtgg ctcacgtgac
1741 catcacgaca aaaacggctg atggaaagtg cgcataccgt gcatcctact cggatgggaa
1801 actacaggcc cctccaaaac cttgtgccgg caaccagggc acactcatca ctgttgaaga
1861 tttgttctac aacattatca cacggcggaa ggctttaaag aatccaagtg aggaatatgg
1921 gaagatcctt gaagtggtcg gacgatactc tatccacaac tctgggattt ccttctctgt
1981 gaagaagcaa ggtgaaactg tttctgatgt gcgcaccttg cccaatgcta ccactgtgga
2041 caatattcgc agcatctttg ggaatgcggt ctctcgagag ctcattgagg ttggatgtga
2101 ggacaagaca ctggctttca agatgaatgg ctacatttcc aatgccaact acagcgttaa
2161 aaagtgtatc ttcctgctgt ttatcaacca taggctggtg gagtcagctg cactccgaaa
2221 agccatagag acagtatatg ctgcatactt gcccaaaaac acgcatccat tcctctacct
2281 gtcgctagaa atctcccctc agaacgtcga tgtcaatgtc caccccacca agcatgaagt
2341 tcacttcctt catgaggaat caatactgca aagagtgcag caacatatag aaagcaaatt
2401 actgggttcc aacagttcaa gaatgtattt cacccagact cttctgcctg gtctggcagg
2461 ccctagtgga gaagcagcac ggccgaccac tggagtggcc tcaagcagta ccagtggttc
2521 tggagacaaa gtgtatgcct accagatggt caggacagac tctcgggagc agaagctcga
2581 cgctttttta cagcctgtgt caagcctggg accctcccag ccacaagatc ctgccccggt
2641 aaggggagcc aggacagaag gctccccaga gcgggccacg agagaggatg aagagatgct
2701 ggcgctccca gcgccagctg aggccgcagc tgaatctgag aatctggaga gagagtctct
2761 gatggagacc tccgatgcag cccagaaagc tgcacccaca tcatctccag gctcttcaag
2821 gaaaaggcac agagaagact ctgatgtaga gatggtcgaa aatgcatcag ggaaagaaat
2881 gacagctgcc tgctacccgc gtaggagaat tataaacctg acaagtgtgt tatcgttgca
2941 ggaggagatt agtgaaaggt gccacgaaac cttgagagag atgctgagga atcatagctt
3001 tgtgggctgt gtgaaccccc aatgggccct ggctcagcac cagaccaaac tatatttgct
3061 taatactacc aagctgtcag aagaactgtt ctatcagata ctcatttatg actttgccaa
3121 ctttggagtt ttgcggctga gcgagcccgc gccactcttt gacctagcca tgcttgctct
3181 ggacagccct gaaagtggct ggactgaaga tgacggcccg aaggaagggt tggctgagta
3241 tattgtagaa tttctgaaga agaaagcaga gatgctcgcc gactacttct ccgtagagat
3301 cgacgaggag ggcaatctta tcggattacc tctcctcata gactcttatg tgcccccttt
3361 ggaggggctg cccattttca tcctccgcct cgctactgaa gtaaactggg atgaggagaa
3421 ggagtgcttc gagagtctta gtaaagagtg tgcaatgttt tatagcatta ggaagcagta
3481 catcctggaa gagtccacac tgtctgggca gcagtcagac atgcctggta gcaccagcaa
3541 gccatggaag tggactgtgg agcacatcat ctacaaggcc ttccggtccc acctgcttcc
3601 tcccaagcac tttaccgagg atgggaatgt gctccagctg gccaatctcc ctgatctgta
3661 caaagttttt taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
3721 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
3781 accctggaag gtgccactcc cactgtcctt tcctaataaa atgagaaaat tgcatcgcat
3841 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
3901 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
3961 gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg
4021 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
4081 ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg
4141 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
4201 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
4261 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
4321 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
4381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
4441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
4501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
4561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
4621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
4681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
4741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
4801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
4861 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
4921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
4981 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
5041 gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
5101 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
5161 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
5221 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
5281 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
5341 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
5401 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
5461 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
5521 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
5581 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
5641 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
5701 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
5761 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
5821 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
5881 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
5941 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
6001 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
6061 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
6121 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
6181 ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat
6241 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
6301 gcgagcaaaa tttaa
//
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