| 质粒名称: | pLV302 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 119943 |
| 存储实验室: | Gerald Schwank |
|
载体骨架基本信息 |
|
| 载体名称: | px601 |
| 载体出品公司: | Feng Zhang lab |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3938 bp |
| 完整质粒大小: | 7283 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,AAV，CRISPR |
| 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: | N-terminal SaKKH-BE3(739) - N-Intein (Npu DnaE)，split BE3 |
| 物种来源: | R. norvegicus (rat); S. aureus |
| 插入大小: | 3345 bp |
| 启动子 | CMV |
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | CGCAAATGGGCGGTAGGCGTG (CMV for) |
| 3' 测序引物 | CCTCGACTGTGCCTTCTA (bGH rev) |
|
文献引用方法 |
|
| 材料和方法部分: | pLV302 was a gift from Gerald Schwank (Addgene plasmid # 119943 ; http://n2t.net/addgene:119943 ; RRID:Addgene_119943) |
| 参考文献部分: | Treatment of a metabolic liver disease by in vivo genome base editing in adult mice. Villiger L, Grisch-Chan HM, Lindsay H, Ringnalda F, Pogliano CB, Allegri G, Fingerhut R, Haberle J, Matos J, Robinson MD, Thony B, Schwank G. Nat Med. 2018 Oct;24(10):1519-1525. doi: 10.1038/s41591-018-0209-1. Epub 2018 Oct 8. 10.1038/s41591-018-0209-1 [pii] PubMed 30297904 |
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 pLV302 7283 bp ds-DNA circular SYN 20-DEC-2018
DEFINITION Expresses N-terminal Npu DnaE Intein-SaKKH-BE3 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pLV302
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7283)
AUTHORS Villiger L, Grisch-Chan HM, Lindsay H, Ringnalda F, Pogliano CB,
Allegri G, Fingerhut R, Haberle J, Matos J, Robinson MD, Thony B,
Schwank G
TITLE Treatment of a metabolic liver disease by in vivo genome base
editing in adult mice.
JOURNAL Nat Med. 2018 Oct;24(10):1519-1525. doi: 10.1038/s41591-018-0209-1.
Epub 2018 Oct 8.
PUBMED 30297904
REFERENCE 2 (bases 1 to 7283)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 20, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7283
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR
enhancer 178..557
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 558..761
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 712..732
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 780..789
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 792..812
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 822..845
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 870..1556
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
primer_bind complement(4149..4166)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4155..4362
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 4371..4511
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4371..4500
/label=AAV2 ITR
rep_origin 4586..5041
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4673..4692)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4883..4904
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5058..5077)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5177..5199
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5237..5255)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5323..5427
/gene="bla"
/label=AmpR promoter
CDS 5428..6288
/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(5646..5665)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6459..7047
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6948..6967
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7201..7218
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctactcg aggcgttgac
181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
421 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
481 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
541 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
661 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actaccggtg
781 ccaccatggg acccaagaag aagaggaaag tcgccgcagc ggactacaag gatgatgatg
841 ataaaggtat tcacggggtc ccagccgcga tgagctcaga gactggccca gtggctgtgg
901 accccacatt gagacggcgg atcgagcccc atgagtttga ggtattcttc gatccgagag
961 agctccgcaa ggagacctgc ctgctttacg aaattaattg ggggggccgg cactccattt
1021 ggcgacatac atcacagaac actaacaagc acgtcgaagt caacttcatc gagaagttca
1081 cgacagaaag atatttctgt ccgaacacaa ggtgcagcat tacctggttt ctcagctgga
1141 gcccatgcgg cgaatgtagt agggccatca ctgaattcct gtcaaggtat ccccacgtca
1201 ctctgtttat ttacatcgca aggctgtacc accacgctga cccccgcaat cgacaaggcc
1261 tgcgggattt gatctcttca ggtgtgacta tccaaattat gactgagcag gagtcaggat
1321 actgctggag aaactttgtg aattatagcc cgagtaatga agcccactgg cctaggtatc
1381 cccatctgtg ggtacgactg tacgttcttg aactgtactg catcatactg ggcctgcctc
1441 cttgtctcaa cattctgaga aggaagcagc cacagctgac attctttacc atcgctcttc
1501 agtcttgtca ttaccagcga ctgcccccac acattctctg ggccaccggg ttgaaaagcg
1561 gcagcgagac tcccgggacc tcagagtccg ccacacccga aagtgggaaa cggaactaca
1621 tcctggggct tgccattggg ataaccagcg ttggctacgg aattattgat tatgagacac
1681 gcgatgtgat tgacgccggg gttaggctgt tcaaagaggc caacgttgaa aacaacgagg
1741 gaagacggag taagcgcgga gcaagaagac tcaagcgcag acggagacat cggattcaga
1801 gggtgaaaaa gctgctcttc gattacaatc tcctgaccga tcatagtgag ctgagcggaa
1861 tcaaccccta cgaggcgcga gtgaaagggc tttcccagaa gctgtccgaa gaggagttct
1921 ccgccgcgtt gctgcacctg gccaaacgga ggggggttca caatgtaaac gaagtggagg
1981 aggacacggg caatgaactt agtacgaaag aacagatcag taggaactct aaggctctcg
2041 aagagaaata cgtcgctgag ttgcagcttg agagactgaa aaaagacggc gaagtacgcg
2101 gatctattaa taggttcaag acttcagatt acgtaaagga agccaagcag ctcctgaaag
2161 tacagaaagc gtaccatcag ctcgatcaga gcttcatcga tacctacata gatttgctgg
2221 agacacggag gacatactac gagggcccag gggaaggatc tccttttggg tggaaggaca
2281 tcaaggaatg gtacgagatg cttatgggac attgtacata ttttccggag gagctcagga
2341 gcgtcaagta cgcctacaat gccgacctgt acaatgccct caatgacctc aataacctcg
2401 tgattaccag ggacgagaac gagaagctgg agtactatga aaagttccag attatcgaga
2461 atgtgtttaa gcagaagaag aagccgacac ttaagcagat tgcaaaggaa atcctcgtga
2521 atgaggaaga tatcaaggga tacagagtga caagtacagg caagcccgag ttcacaaatc
2581 tgaaggtgta ccacgatatt aaggacataa ccgcacgaaa ggagataatc gaaaacgctg
2641 agctcctcga tcagatcgca aaaattctta ccatctacca gtctagtgag gacattcagg
2701 aggaactgac taatctgaac agtgagctca cccaagagga aattgagcag atttcaaacc
2761 tgaaaggcta caccgggacg cacaatctga gcctcaaagc aatcaacctc attctggatg
2821 aactttggca cacaaatgac aaccaaattg ccatattcaa ccgcctgaaa ctggtgccaa
2881 aaaaagtgga tctgtcacag caaaaggaaa tccctacaac cttggttgac gattttattc
2941 tgtcccccgt tgtcaagcgg agcttcatcc agtcaatcaa ggtgatcaat gccatcatta
3001 aaaaatacgg attgccaaac gatataatta tcgagcttgc acgagagaag aactcaaagg
3061 acgcccagaa gatgattaac gaaatgcaga agcgcaaccg ccagacaaac gaacgcatag
3121 aggaaattat aagaacaacc ggcaaagaga atgccaagta tctgatcgag aaaatcaagc
3181 tgcacgacat gcaagaaggc aagtgcctgt actctctgga agctatccca ctcgaagatc
3241 tgctgaataa tccattcaat tacgaggtgg accacatcat ccctagatcc gtaagctttg
3301 acaattcctt caataacaaa gttctggtta aacaggagga aaattctaaa aaagggaacc
3361 ggaccccgtt ccagtacctg agctccagtg acagcaagat tagctacgag acttttaaga
3421 aacatattct gaatctggcc aaaggcaaag gcaggatcag caagaccaag aaggagtacc
3481 tcctcgaaga acgcgacatt aacagattta gtgtgcagaa agatttcatc aaccgaaacc
3541 ttgtcgatac tcggtacgcc acgagaggcc tgatgaatct cctcaggagc tacttccgcg
3601 tcaataatct ggacgttaaa gtcaagagca taaatggggg attcaccagc tttctgagga
3661 gaaagtggaa gtttaagaag gaacgaaaca aaggatacaa gcaccatgct gaggatgctt
3721 tgatcatcgc taacgcggac tttatcttta aggaatggaa aaagctggat aaggcaaaga
3781 aagtgatgga aaaccagatg ttcgaggaga agcaggcaga gtgtcttagc tacgagacag
3841 agattcttac agtggagtat ggacttcttc ccatcggtaa aattgttgaa aagaggatcg
3901 agtgcacggt ctacagtgtc gataataatg gtaacatata tacgcaaccc gtcgcgcagt
3961 ggcatgatag aggcgagcag gaagtatttg agtattgtct ggaggatgga tctctgattc
4021 gcgcgactaa ggaccataag tttatgacgg ttgacggaca aatgctccca atagacgaga
4081 tatttgaacg agaactggat cttatgagag tcgataactt gccgaattaa ctagagctcg
4141 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
4201 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
4261 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
4321 caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct
4381 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
4441 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
4501 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
4561 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
4621 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
4681 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
4741 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
4801 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
4861 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
4921 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
4981 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
5041 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
5101 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
5161 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
5221 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
5281 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
5341 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
5401 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
5461 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
5521 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
5581 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
5641 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
5701 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
5761 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
5821 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
5881 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
5941 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
6001 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
6061 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
6121 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
6181 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
6241 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
6301 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
6361 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
6421 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
6481 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
6541 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
6601 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
6661 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
6721 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
6781 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
6841 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
6901 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
6961 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
7021 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
7081 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
7141 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
7201 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
7261 cgttggccga ttcattaatg cag
//
