| 质粒名称: | SaBE4 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 100805 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3402 bp |
| 完整质粒大小: | 8034 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 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: | SaBE4 |
| 物种来源: | R、 褐家鼠，合成；金黄色葡萄球菌 |
| 插入大小: | 4632 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 吉布森克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 |
| 3' 测序引物 | M13R |
|
文献引用方法 |
|
| 材料和方法部分: | SaBE4 was a gift from David Liu (Addgene plasmid # 100805 ; http://n2t.net/addgene:100805 ; RRID:Addgene_100805) |
| 参考文献部分: | Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and product purity. Komor AC, Zhao KT, Packer MS, Gaudelli NM, Waterbury AL, Koblan LW, Kim YB, Badran AH, Liu DR. Sci Adv. 2017 Aug 30;3(8):eaao4774. doi: 10.1126/sciadv.aao4774. eCollection 2017 Aug. aao4774 [pii] PubMed 28875174 |
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 SaBE4 8034 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses SaBE4 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS SaBE4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8034)
AUTHORS Komor AC, Zhao KT, Packer MS, Gaudelli NM, Waterbury AL, Koblan LW,
Kim YB, Badran AH, Liu DR
TITLE Improved base excision repair inhibition and bacteriophage Mu Gam
protein yields C:G-to-T:A base editors with higher efficiency and
product purity.
JOURNAL Sci Adv. 2017 Aug 30;3(8):eaao4774. doi: 10.1126/sciadv.aao4774.
eCollection 2017 Aug.
PUBMED 28875174
REFERENCE 2 (bases 1 to 8034)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8034
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 132..335
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 286..306
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 377..396
/label=T7
/note="T7 promoter, forward primer"
promoter 377..395
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 409..1095
/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"
CDS 4357..4377
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4384..4449
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 4480..4728
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 4759..5007
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5020..5040
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5049..5066
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5089..5106)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5095..5319
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5390..5406)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5390..5406)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5403..5425)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5414..5430
/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(5438..5468)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5483..5504
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5621..5638)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5792..6380)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5872..5891)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6551..7411)
/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 7174..7193
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7412..7516)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7595..7614)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 7786..131
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gagctcagag
421 actggcccag tggctgtgga ccccacattg agacggcgga tcgagcccca tgagtttgag
481 gtattcttcg atccgagaga gctccgcaag gagacctgcc tgctttacga aattaattgg
541 gggggccggc actccatttg gcgacataca tcacagaaca ctaacaagca cgtcgaagtc
601 aacttcatcg agaagttcac gacagaaaga tatttctgtc cgaacacaag gtgcagcatt
661 acctggtttc tcagctggag cccatgcggc gaatgtagta gggccatcac tgaattcctg
721 tcaaggtatc cccacgtcac tctgtttatt tacatcgcaa ggctgtacca ccacgctgac
781 ccccgcaatc gacaaggcct gcgggatttg atctcttcag gtgtgactat ccaaattatg
841 actgagcagg agtcaggata ctgctggaga aactttgtga attatagccc gagtaatgaa
901 gcccactggc ctaggtatcc ccatctgtgg gtacgactgt acgttcttga actgtactgc
961 atcatactgg gcctgcctcc ttgtctcaac attctgagaa ggaagcagcc acagctgaca
1021 ttctttacca tcgctcttca gtcttgtcat taccagcgac tgcccccaca cattctctgg
1081 gccaccgggt tgaaatctgg tggttcttct ggtggttcta gcggcagcga gactcccggg
1141 acctcagagt ccgccacacc cgaaagttct ggtggttctt ctggtggttc tgggaaacgg
1201 aactacatcc tggggcttgc cattgggata accagcgttg gctacggaat tattgattat
1261 gagacacgcg atgtgattga cgccggggtt aggctgttca aagaggccaa cgttgaaaac
1321 aacgagggaa gacggagtaa gcgcggagca agaagactca agcgcagacg gagacatcgg
1381 attcagaggg tgaaaaagct gctcttcgat tacaatctcc tgaccgatca tagtgagctg
1441 agcggaatca acccctacga ggcgcgagtg aaagggcttt cccagaagct gtccgaagag
1501 gagttctccg ccgcgttgct gcacctggcc aaacggaggg gggttcacaa tgtaaacgaa
1561 gtggaggagg acacgggcaa tgaacttagt acgaaagaac agatcagtag gaactctaag
1621 gctctcgaag agaaatacgt cgctgagttg cagcttgaga gactgaaaaa agacggcgaa
1681 gtacgcggat ctattaatag gttcaagact tcagattacg taaaggaagc caagcagctc
1741 ctgaaagtac agaaagcgta ccatcagctc gatcagagct tcatcgatac ctacatagat
1801 ttgctggaga cacggaggac atactacgag ggcccagggg aaggatctcc ttttgggtgg
1861 aaggacatca aggaatggta cgagatgctt atgggacatt gtacatattt tccggaggag
1921 ctcaggagcg tcaagtacgc ctacaatgcc gacctgtaca atgccctcaa tgacctcaat
1981 aacctcgtga ttaccaggga cgagaacgag aagctggagt actatgaaaa gttccagatt
2041 atcgagaatg tgtttaagca gaagaagaag ccgacactta agcagattgc aaaggaaatc
2101 ctcgtgaatg aggaagatat caagggatac agagtgacaa gtacaggcaa gcccgagttc
2161 acaaatctga aggtgtacca cgatattaag gacataaccg cacgaaagga gataatcgaa
2221 aacgctgagc tcctcgatca gatcgcaaaa attcttacca tctaccagtc tagtgaggac
2281 attcaggagg aactgactaa tctgaacagt gagctcaccc aagaggaaat tgagcagatt
2341 tcaaacctga aaggctacac cgggacgcac aatctgagcc tcaaagcaat caacctcatt
2401 ctggatgaac tttggcacac aaatgacaac caaattgcca tattcaaccg cctgaaactg
2461 gtgccaaaaa aagtggatct gtcacagcaa aaggaaatcc ctacaacctt ggttgacgat
2521 tttattctgt cccccgttgt caagcggagc ttcatccagt caatcaaggt gatcaatgcc
2581 atcattaaaa aatacggatt gccaaacgat ataattatcg agcttgcacg agagaagaac
2641 tcaaaggacg cccagaagat gattaacgaa atgcagaagc gcaaccgcca gacaaacgaa
2701 cgcatagagg aaattataag aacaaccggc aaagagaatg ccaagtatct gatcgagaaa
2761 atcaagctgc acgacatgca agaaggcaag tgcctgtact ctctggaagc tatcccactc
2821 gaagatctgc tgaataatcc attcaattac gaggtggacc acatcatccc tagatccgta
2881 agctttgaca attccttcaa taacaaagtt ctggttaaac aggaggaaaa ttctaaaaaa
2941 gggaaccgga ccccgttcca gtacctgagc tccagtgaca gcaagattag ctacgagact
3001 tttaagaaac atattctgaa tctggccaaa ggcaaaggca ggatcagcaa gaccaagaag
3061 gagtacctcc tcgaagaacg cgacattaac agatttagtg tgcagaaaga tttcatcaac
3121 cgaaaccttg tcgatactcg gtacgccacg agaggcctga tgaatctcct caggagctac
3181 ttccgcgtca ataatctgga cgttaaagtc aagagcataa atgggggatt caccagcttt
3241 ctgaggagaa agtggaagtt taagaaggaa cgaaacaaag gatacaagca ccatgctgag
3301 gatgctttga tcatcgctaa cgcggacttt atctttaagg aatggaaaaa gctggataag
3361 gcaaagaaag tgatggaaaa ccagatgttc gaggagaagc aggcagagtc aatgcctgag
3421 atcgagacag agcaggaata caaggaaatt ttcatcaccc ctcatcagat taaacacata
3481 aaggacttca aagactataa atactctcat agggtggaca aaaaacccaa tcgcgagctc
3541 attaatgaca ccctgtactc aacacggaag gatgataaag gtaatacctt gattgtgaat
3601 aatcttaatg gattgtatga caaagataac gacaagctca agaagctgat caacaagtct
3661 ccagagaagc tccttatgta tcaccacgac ccacagactt atcagaaatt gaaactgatc
3721 atggagcaat acggggatga gaagaaccca ctctacaaat attatgagga aacaggtaat
3781 tacctgacca agtactccaa gaaggataac ggaccagtga tcaaaaagat aaagtactat
3841 ggcaacaaac ttaatgcgca tttggacata actgacgatt accccaattc tcgaaacaag
3901 gttgtgaagc tctccctgaa gccttataga tttgacgtgt acctggataa tggggtttat
3961 aaattcgtca ccgtgaaaaa tctggacgtg atcaaaaagg agaactatta tgaagtaaac
4021 tcaaagtgct atgaggaggc gaagaagctg aagaagatct ccaatcaggc cgagttcatc
4081 gcttccttct ataataacga tctcatcaag atcaatggag agctttatcg cgtcattggt
4141 gtgaacaatg acttgctgaa caggatcgaa gtcaatatga tagacattac ctaccgggag
4201 tatctcgaaa acatgaatga taaacggccg cctcgcatca tcaagacaat cgcatctaaa
4261 actcagtcaa taaaaaagta ctctaccgat atcctgggga atctctatga agtgaagtca
4321 aagaagcacc cacaaatcat taaaaaaggt ggatccccca agaagaagag gaaagtctcg
4381 agcgactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
4441 gatgacaagt ctggtggttc tggaggatct ggtggttcta ctaatctgtc agatattatt
4501 gaaaaggaga ccggtaagca actggttatc caggaatcca tcctcatgct cccagaggag
4561 gtggaagaag tcattgggaa caagccggaa agcgatatac tcgtgcacac cgcctacgac
4621 gagagcaccg acgagaatgt catgcttctg actagcgacg cccctgaata caagccttgg
4681 gctctggtca tacaggatag caacggtgag aacaagatta agatgctctc tggtggttct
4741 ggaggatctg gtggttctac taatctgtca gatattattg aaaaggagac cggtaagcaa
4801 ctggttatcc aggaatccat cctcatgctc ccagaggagg tggaagaagt cattgggaac
4861 aagccggaaa gcgatatact cgtgcacacc gcctacgacg agagcaccga cgagaatgtc
4921 atgcttctga ctagcgacgc ccctgaatac aagccttggg ctctggtcat acaggatagc
4981 aacggtgaga acaagattaa gatgctctct ggtggttctc ccaagaagaa gaggaaagtc
5041 taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
5101 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
5161 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
5221 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
5281 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
5341 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
5401 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
5461 agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
5521 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
5581 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
5641 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
5701 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
5761 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
5821 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
5881 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
5941 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
6001 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
6061 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
6121 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
6181 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
6241 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
6301 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
6361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
6421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
6481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
6541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
6601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
6661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
6721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
6781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
6841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
6901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
6961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
7021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
7081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
7141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
7201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
7261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
7321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
7381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
7441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
7501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
7561 atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt
7621 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
7681 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
7741 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
7801 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
7861 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
7921 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
7981 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatc
//
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