| 质粒名称: | Cbh_v5 AAV-CBE C-terminal |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 137176 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | px601 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | AAV |
| 筛选标记: | -- |
| 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: | v5 AAV-CBE C-terminal; U6-protospacer，JL628 |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Cbh |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | Cbh_v5 AAV-CBE C-terminal was a gift from David Liu (Addgene plasmid # 137176 ; http://n2t.net/addgene:137176 ; RRID:Addgene_137176) |
| 参考文献部分: | Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated viruses. Levy JM, Yeh WH, Pendse N, Davis JR, Hennessey E, Butcher R, Koblan LW, Comander J, Liu Q, Liu DR. Nat Biomed Eng. 2020 Jan;4(1):97-110. doi: 10.1038/s41551-019-0501-5. Epub 2020 Jan 14. 10.1038/s41551-019-0501-5 PubMed 31937940 |
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 Cbh_v5 AAV-CBE C-terminal 7658 bp ds-DNA circular SYN 28-JAN-2020
DEFINITION AAV genome: expresses the C-terminal of v5 AAV-CBE from the Cbh
promoter, and U6-sgRNA.
ACCESSION .
VERSION .
KEYWORDS Cbh_v5 AAV-CBE C-terminal
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7658)
AUTHORS Levy JM, Yeh WH, Pendse N, Davis JR, Hennessey E, Butcher R, Koblan
LW, Comander J, Liu Q, Liu DR
TITLE Cytosine and adenine base editing of the brain, liver, retina, heart
and skeletal muscle of mice via adeno-associated viruses.
JOURNAL Nat Biomed Eng. 2020 Jan;4(1):97-110. doi:
10.1038/s41551-019-0501-5. Epub 2020 Jan 14.
PUBMED 31937940
REFERENCE 2 (bases 1 to 7658)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 28, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7658
/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..3633
/label=chicken beta-actin promoter
intron 3634..3861
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 3915..3935
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4041..6425
/codon_start=1
/product="C-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(C)
/translation="CFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILED
IVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK
TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKG
ILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQI
LKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDN
KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDK
AGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGK
ATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMP
QVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKV
EKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELEN
GRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLD
EIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY
FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
CDS 6456..6704
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 6747..6767
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(6832..6852)
/label=WPRE-R
/note="WPRE, reverse primer"
polyA_signal 7028..7252
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_RNA complement(7266..7341)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(7371..7611)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(7421..7440)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(7591..7611)
/label=hU6-F
/note="Human U6 promoter, forward primer"
repeat_region 7635..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 gggggggsgc
3481 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
3541 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
3601 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
3661 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
3721 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
3781 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
3841 cctgaaatca ctttttttca ggttggaccg gtgccaccat gaaacggaca gccgacggaa
3901 gcgagttcga gtcaccaaag aagaagcgga aagtcatcaa gattgctaca cggaaatacc
3961 tgggaaagca gaacgtgtac gacatcggcg tggagcggga tcacaacttc gccctgaaga
4021 atggctttat cgccagcaat tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
4081 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
4141 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
4201 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
4261 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
4321 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
4381 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
4441 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
4501 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
4561 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
4621 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
4681 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
4741 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
4801 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
4861 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
4921 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
4981 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
5041 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
5101 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
5161 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
5221 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
5281 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
5341 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
5401 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
5461 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
5521 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
5581 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
5641 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
5701 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
5761 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
5821 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
5881 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
5941 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
6001 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
6061 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
6121 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
6181 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
6241 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
6301 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
6361 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
6421 gtgacagcgg cgggagcggc gggagcgggg ggagcactaa tctgagcgac atcattgaga
6481 aggagactgg gaaacagctg gtcattcagg agtccatcct gatgctgcct gaggaggtgg
6541 aggaagtgat cggcaacaag ccagagtctg acatcctggt gcacaccgcc tacgacgagt
6601 ccacagatga gaatgtgatg ctgctgacct ctgacgcccc cgagtataag ccttgggccc
6661 tggtcatcca ggattctaac ggcgagaata agatcaagat gctgagcgga ggatccaaaa
6721 gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtctaa gatctgataa
6781 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
6841 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
6901 ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat
6961 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
7021 ggtgcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
7081 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
7141 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
7201 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggctcgagaa
7261 aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct tattttaact
7321 tgctatttct agctctaaaa ccgagacgca ttaatcgtct cgggtgtttc gtcctttcca
7381 caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca tatgatagtc
7441 cattttaaaa cataatttta aaactgcaaa ctacccaaga aattattact ttctacgtca
7501 cgtattttgt actaatatct ttgtgtttac agtcaaatta attctaatta tctctctaac
7561 agccttgtat cgtatatgca aatatgaagg aatcatggga aataggccct cttcctgccc
7621 gaccttgcgg ccgcaggaac ccctagtgat ggagttgg
//
