| 质粒名称: | v3em-Nterm-PE2max |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 198734 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | p601 modified |
| 空载体大小: | 4739 bp |
| 完整质粒大小: | 7549bp |
| 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: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | Nterm PE2max-NpuN |
| 插入大小: | 3500 bp |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | AGCGAGTCAGTGAGCGAG |
| 3’测序引物 | GTTTTCCCAGTCACGAC |
|
文献引用方法 |
|
| 材料和方法部分: | v3em-Nterm-PE2max was a gift from David Liu (Addgene plasmid # 198734 ; http://n2t.net/addgene:198734 ; RRID:Addgene_198734) |
| 参考文献部分: | Efficient prime editing in mouse brain, liver and heart with dual AAVs. Davis JR, Banskota S, Levy JM, Newby GA, Wang X, Anzalone AV, Nelson AT, Chen PJ, Hennes AD, An M, Roh H, Randolph PB, Musunuru K, Liu DR. Nat Biotechnol. 2023 May 4. doi: 10.1038/s41587-023-01758-z. 10.1038/s41587-023-01758-z PubMed 37142705 |
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 v3em-Nterm-PE2max 7549 bp ds-DNA circular SYN 19-APR-2023
DEFINITION AAV genome encoding N-terminal PE2max.
ACCESSION .
VERSION .
KEYWORDS v3em-Nterm-PE2max
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7549)
TITLE Efficient AAV-Mediated In Vivo Prime Editing in Multiple Organs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7549)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 19, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7549
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(125..142)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(125..141)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(134..156)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 283..738
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(370..389)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 580..601
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 764..868
/gene="bla"
/label=AmpR promoter
CDS 869..1729
/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 complement(1087..1106)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1900..2488
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2389..2408
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2642..2659
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2776..2797
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2812..2842
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2850..2866
/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)."
primer_bind 2855..2877
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2874..2890
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2874..2890
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
repeat_region 2917..3046
/label=AAV2 ITR
enhancer 3069..3354
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 3356..3632
/label=chicken beta-actin promoter
intron 3633..3860
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 3914..3934
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3935..5650
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
CDS 7355..7375
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind 7395..7414
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
repeat_region 7526..106
/label=AAV2 ITR
ORIGIN
1 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
61 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta
121 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
181 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
241 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
481 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
541 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
601 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
661 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
721 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
781 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
841 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
901 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
961 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
1021 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
1081 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
1141 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
1201 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
1261 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
1321 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
1381 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
1441 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
1501 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
1561 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
1621 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
1681 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
1741 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
1801 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
1861 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
1921 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
1981 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
2041 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
2101 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
2161 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
2221 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
2281 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
2341 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
2401 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
2461 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
2521 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
2581 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
2641 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
2701 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
2761 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
2821 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
2881 cagctatgac catgattacg ccagatttaa ttaaggctgc gcgctcgctc gctcactgag
2941 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
3001 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg cctctagatc
3061 agggtacccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
3121 cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag
3181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
3241 cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta
3301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
3361 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
3421 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg ggggggcgcg
3481 cgccaggggg gggggggggg ggsgaggggg ggggsggggs gaggsggaga ggtgcggcgg
3541 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
3601 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
3661 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
3721 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt
3781 aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc
3841 ctgaaatcac tttttttcag gttggaccgg tgccaccatg aaacggacag ccgacggaag
3901 cgagttcgag tcaccaaaga agaagcggaa agtcgacaag aagtacagca tcggcctgga
3961 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
4021 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
4081 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
4141 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
4201 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
4261 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
4321 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
4381 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
4441 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
4501 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
4561 catcctgtct gccagactga gcaagagcag aaagctggaa aatctgatcg cccagctgcc
4621 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
4681 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
4741 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
4801 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
4861 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
4921 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
4981 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
5041 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
5101 cgtgaagctg aagagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
5161 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
5221 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
5281 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
5341 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
5401 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
5461 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
5521 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
5581 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
5641 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
5701 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
5761 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
5821 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
5881 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
5941 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
6001 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
6061 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
6121 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
6181 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
6241 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
6301 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
6361 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
6421 caaccggctg tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga
6481 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
6541 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
6601 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
6661 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
6721 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
6781 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
6841 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
6901 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
6961 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagtgcctgt cctacgagac
7021 agagatcctg acagtggagt atggcctgct gccaatcggc aagatcgtgg agaagaggat
7081 cgagtgtacc gtgtactctg tggataacaa tggcaacatc tatacacagc ccgtggcaca
7141 gtggcacgat aggggagagc aggaggtgtt cgagtattgc ctggaggacg gcagcctgat
7201 cagggcaacc aaggaccaca agttcatgac agtggatggc cagatgctgc ccatcgacga
7261 gattttcgag cgggagctgg acctgatgag agtggataac ctgcctaatt ctggcggctc
7321 aaaaagaacc gccgacggca gcgaattcga gtctcccaag aagaagagga aagtctaagt
7381 cgacctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
7441 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
7501 gggaggtttt ttaaagcgcg gccgcaggaa cccctagtga tggagttgg
//
