| 质粒名称: | pT7-hMLH1dn for IVT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 178114 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pT7 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 5765bp |
| 载体类型: | Template for in vitro transcription |
| 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: | human MLH1dn (codon optimized) // homo sapiens MLH1 del754-756 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2259bp |
| 基因突变: | Detailed in manuscript |
| 启动子: | T7 (inactivated) |
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pT7-hMLH1dn for IVT was a gift from David Liu (Addgene plasmid # 178114 ; http://n2t.net/addgene:178114 ; RRID:Addgene_178114) |
| 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 |
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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 pT7-hMLH1dn for IVT 5765 bp ds-DNA circular SYN 30-NOV-2021
DEFINITION Template for in vitro transcription of human MLH1dn.
ACCESSION .
VERSION .
KEYWORDS pT7-hMLH1dn for IVT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5765)
AUTHORS 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
TITLE Enhanced prime editing systems by manipulating cellular determinants
of editing outcomes.
JOURNAL Cell. 2021 Oct 9. pii: S0092-8674(21)01065-5. doi:
10.1016/j.cell.2021.09.018.
PUBMED 34653350
REFERENCE 2 (bases 1 to 5765)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 30, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5765
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(498..1358)
/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 1121..1140
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1359..1463)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1542..1561)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 1733..2112
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2113..2316
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2267..2287
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind complement(4801..4818)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4807..5031
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5102..5118)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5102..5118)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5115..5137)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5126..5142
/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(5150..5180)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5195..5216
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5333..5350)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5504..327)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5584..5603)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
61 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
361 actcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
1021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
1081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
1141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
1201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
1261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
1321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
1381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
1441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
1501 tcgggagatc gatctcccga tcccctaggg tctactctca gtacaatctg ctctgatgcc
1561 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
1621 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
1681 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
1741 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
1801 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
1861 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
1921 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
1981 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
2041 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
2101 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
2161 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
2221 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
2281 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatctcg
2341 agctcggtac ctaatacgac acactataag gaaataagag agaaaagaag agtaagaaga
2401 aatataagag ccaccatgag cttcgttgct ggagtcatcc ggagactgga cgagacagtg
2461 gtgaacagaa ttgccgccgg cgaggtgatc cagagacctg ccaatgcaat aaaggagatg
2521 atcgagaact gtctggacgc caagtccaca agcattcagg tgatcgtgaa ggagggcgga
2581 ctgaagctga tccagatcca agacaacggc acaggcatca gaaaggaaga tctggacatc
2641 gtgtgtgaac ggttcaccac atctaagctg cagtcttttg aggatctggc ctctatcagt
2701 acctacggct tcagaggcga ggccctggcc agcatcagcc acgtggccca tgtgaccatc
2761 accaccaaaa ccgccgacgg caaatgcgct tatcgcgcta gctacagcga cggcaagctg
2821 aaagccccgc caaagccttg cgccggcaac cagggtacac agataacagt ggaggatctg
2881 ttctacaaca tcgccacccg gagaaaggcc ctgaaaaatc ccagcgagga gtacggcaag
2941 atcctggaag tcgtcggcag atactccgtg cacaacgccg gaatcagctt tagcgtaaag
3001 aagcagggag aaaccgtggc cgatgtgcgc accctgccaa atgccagcac cgtggataac
3061 atcagaagca ttttcggaaa tgccgtgtcc agagaactga tcgagatcgg ctgcgaagat
3121 aagaccctgg cttttaagat gaacggctac atcagcaacg ccaattactc tgtgaagaag
3181 tgcatctttc ttctgttcat caaccacaga ctggtggaaa gcaccagcct gcggaaagcc
3241 atcgagacag tgtacgccgc ctacctgcct aagaacaccc accccttcct gtacctgagc
3301 ctcgagatca gccctcagaa cgtggacgtc aatgtgcatc ctacaaagca cgaggtgcac
3361 ttcctgcacg aggaaagcat cctggaaaga gtgcagcagc acattgagag caagctgctg
3421 ggctctaaca gcagcagaat gtacttcaca cagaccctgc tgcctggcct ggccggcccc
3481 tcaggcgaaa tggttaagtc cacaacctct ctgacctcat ccagcaccag cggttcttcc
3541 gataaggtgt acgcccacca gatggtgcgg accgactctc gggagcagaa gctggacgcc
3601 tttctgcaac ctctgagcaa acctctgagc tctcagcctc aggccatcgt gaccgaggac
3661 aagacagata tctcctccgg ccgtgccaga cagcaggacg aagaaatgct cgagctgcca
3721 gctcctgccg aggtggccgc caagaaccag agcctggagg gagataccac aaagggcacc
3781 agcgaaatga gcgagaagcg gggccctacc tccagcaacc ccagaaaacg gcaccgggag
3841 gacagcgacg tggaaatggt ggaggacgac agcagaaagg aaatgacagc cgcttgtacc
3901 cctagaagaa gaatcatcaa cctgacctcc gtgctgagcc tgcaggagga gatcaacgag
3961 cagggccacg aggtgctgag agagatgctg cacaatcaca gcttcgtggg ctgcgtgaac
4021 cctcaatggg ccctggctca gcatcaaaca aagctgtacc tgctgaacac caccaagctg
4081 agcgaagagc tgttctacca gatcctcatc tacgacttcg ccaacttcgg cgtgctacgc
4141 ctgagcgagc ccgcccctct gtttgacctg gccatgctgg ctctggatag cccagaaagc
4201 ggctggacag aagaggacgg acctaaagag gggctggctg aatacatcgt ggagttcctg
4261 aagaaaaagg ccgagatgct ggccgactac ttttctctgg aaatcgacga ggaaggcaac
4321 ctgatcggcc tgcctctgct gatcgataac tacgtgcctc ccctggaagg cctgcccatc
4381 ttcatcctga gactggctac agaggtgaac tgggacgagg aaaaggaatg cttcgagtct
4441 ctgagcaagg agtgcgccat gttctatagc atcagaaaac agtacatctc tgaagagagc
4501 actctgtctg gccagcagag tgaagtgccc ggaagcatcc ccaacagctg gaagtggacc
4561 gtggaacaca tcgtgtacaa ggccctgcgg agccacattc tccctcctaa gcacttcacc
4621 gaggacggca acatcctgca gctggccaac ctgcccgacc tttataaggt tttctaatta
4681 attaagctgc cttctgcggg gcttgccttc tggccatgcc cttcttctct cccttgcacc
4741 tgtacctctt ggtctttgaa taaagcctga gtaggaagtg agtttaaacc cgctgatcag
4801 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
4861 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgagaaa attgcatcgc
4921 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
4981 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
5041 cggaaagaac cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat
5101 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
5161 ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg
5221 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
5281 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
5341 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
5401 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
5461 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
5521 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
5581 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
5641 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
5701 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
5761 acgaa
//
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