| 质粒名称: | SaBE4-Gam |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 100809 |
| 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: | 8601 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-Gam |
| 物种来源: | R. norvegicus (rat), Synthetic; S. Aureus |
| 插入大小: | 5199 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 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 | T7 |
| 3' 测序引物 | M13R |
|
文献引用方法 |
|
| 材料和方法部分: | SaBE4-Gam was a gift from David Liu (Addgene plasmid # 100809 ; http://n2t.net/addgene:100809 ; RRID:Addgene_100809) |
| 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
SaBE4Gam载体基本信息
SaBE4Gam载体图谱质粒图谱和多克隆位点信息
SaBE4Gam载体简介
SaBE4Gam载体序列
LOCUS SaBE4-Gam 8601 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses SaBE4-Gam in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS SaBE4-Gam
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8601)
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 8601)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8601
/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"
regulatory 403..412
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 979..1662
/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="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
WATGLK"
CDS 4924..4944
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4951..5016
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 5047..5295
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5326..5574
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5587..5607
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5616..5633
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5656..5673)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5662..5886
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5957..5973)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5957..5973)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5970..5992)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5981..5997
/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(6005..6035)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6050..6071
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6188..6205)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6359..6947)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6439..6458)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7118..7978)
/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 7741..7760
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7979..8083)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8162..8181)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 8353..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 ggctaaacca
421 gcaaaacgta tcaagagtgc cgcagcggct tatgtgccac aaaaccgcga tgcggtgatt
481 accgatatta aacgcatcgg ggatttacag cgcgaagcat cacgtctgga aacggaaatg
541 aatgatgcca tcgcggaaat tacggagaaa tttgcggccc ggattgcacc gattaaaacc
601 gatattgaaa ccctttcaaa aggcgttcag ggatggtgtg aagcgaaccg cgacgaactg
661 acgaacggcg gcaaagtgaa gacggcgaat cttgtcaccg gtgatgtatc gtggcgggtc
721 cgtccaccat cagtaagtat tcgtggtatg gatgcagtga tggaaacgct ggagcgtctt
781 ggcctgcaac gctttattcg cacgaagcag gaaatcaaca aggaagcgat tttactggaa
841 ccgaaagcgg tcgcaggcgt tgccggaatt acagttaaat caggcattga ggatttttct
901 attattccat ttgaacagga agccggtatt agcggcagcg agactcccgg gacctcagag
961 tccgctacac ccgaaagtag ctcagagact ggcccagtgg ctgtggaccc cacattgaga
1021 cggcggatcg agccccatga gtttgaggta ttcttcgatc cgagagagct ccgcaaggag
1081 acctgcctgc tttacgaaat taattggggg ggccggcact ccatttggcg acatacatca
1141 cagaacacta acaagcacgt cgaagtcaac ttcatcgaga agttcacgac agaaagatat
1201 ttctgtccga acacaaggtg cagcattacc tggtttctca gctggagccc atgcggcgaa
1261 tgtagtaggg ccatcactga attcctgtca aggtatcccc acgtcactct gtttatttac
1321 atcgcaaggc tgtaccacca cgctgacccc cgcaatcgac aaggcctgcg ggatttgatc
1381 tcttcaggtg tgactatcca aattatgact gagcaggagt caggatactg ctggagaaac
1441 tttgtgaatt atagcccgag taatgaagcc cactggccta ggtatcccca tctgtgggta
1501 cgactgtacg ttcttgaact gtactgcatc atactgggcc tgcctccttg tctcaacatt
1561 ctgagaagga agcagccaca gctgacattc tttaccatcg ctcttcagtc ttgtcattac
1621 cagcgactgc ccccacacat tctctgggcc accgggttga aatctggtgg ttcttctggt
1681 ggttctagcg gcagcgagac tcccgggacc tcagagtccg ccacacccga aagttctggt
1741 ggttcttctg gtggttctgg gaaacggaac tacatcctgg ggcttgccat tgggataacc
1801 agcgttggct acggaattat tgattatgag acacgcgatg tgattgacgc cggggttagg
1861 ctgttcaaag aggccaacgt tgaaaacaac gagggaagac ggagtaagcg cggagcaaga
1921 agactcaagc gcagacggag acatcggatt cagagggtga aaaagctgct cttcgattac
1981 aatctcctga ccgatcatag tgagctgagc ggaatcaacc cctacgaggc gcgagtgaaa
2041 gggctttccc agaagctgtc cgaagaggag ttctccgccg cgttgctgca cctggccaaa
2101 cggagggggg ttcacaatgt aaacgaagtg gaggaggaca cgggcaatga acttagtacg
2161 aaagaacaga tcagtaggaa ctctaaggct ctcgaagaga aatacgtcgc tgagttgcag
2221 cttgagagac tgaaaaaaga cggcgaagta cgcggatcta ttaataggtt caagacttca
2281 gattacgtaa aggaagccaa gcagctcctg aaagtacaga aagcgtacca tcagctcgat
2341 cagagcttca tcgataccta catagatttg ctggagacac ggaggacata ctacgagggc
2401 ccaggggaag gatctccttt tgggtggaag gacatcaagg aatggtacga gatgcttatg
2461 ggacattgta catattttcc ggaggagctc aggagcgtca agtacgccta caatgccgac
2521 ctgtacaatg ccctcaatga cctcaataac ctcgtgatta ccagggacga gaacgagaag
2581 ctggagtact atgaaaagtt ccagattatc gagaatgtgt ttaagcagaa gaagaagccg
2641 acacttaagc agattgcaaa ggaaatcctc gtgaatgagg aagatatcaa gggatacaga
2701 gtgacaagta caggcaagcc cgagttcaca aatctgaagg tgtaccacga tattaaggac
2761 ataaccgcac gaaaggagat aatcgaaaac gctgagctcc tcgatcagat cgcaaaaatt
2821 cttaccatct accagtctag tgaggacatt caggaggaac tgactaatct gaacagtgag
2881 ctcacccaag aggaaattga gcagatttca aacctgaaag gctacaccgg gacgcacaat
2941 ctgagcctca aagcaatcaa cctcattctg gatgaacttt ggcacacaaa tgacaaccaa
3001 attgccatat tcaaccgcct gaaactggtg ccaaaaaaag tggatctgtc acagcaaaag
3061 gaaatcccta caaccttggt tgacgatttt attctgtccc ccgttgtcaa gcggagcttc
3121 atccagtcaa tcaaggtgat caatgccatc attaaaaaat acggattgcc aaacgatata
3181 attatcgagc ttgcacgaga gaagaactca aaggacgccc agaagatgat taacgaaatg
3241 cagaagcgca accgccagac aaacgaacgc atagaggaaa ttataagaac aaccggcaaa
3301 gagaatgcca agtatctgat cgagaaaatc aagctgcacg acatgcaaga aggcaagtgc
3361 ctgtactctc tggaagctat cccactcgaa gatctgctga ataatccatt caattacgag
3421 gtggaccaca tcatccctag atccgtaagc tttgacaatt ccttcaataa caaagttctg
3481 gttaaacagg aggaaaattc taaaaaaggg aaccggaccc cgttccagta cctgagctcc
3541 agtgacagca agattagcta cgagactttt aagaaacata ttctgaatct ggccaaaggc
3601 aaaggcagga tcagcaagac caagaaggag tacctcctcg aagaacgcga cattaacaga
3661 tttagtgtgc agaaagattt catcaaccga aaccttgtcg atactcggta cgccacgaga
3721 ggcctgatga atctcctcag gagctacttc cgcgtcaata atctggacgt taaagtcaag
3781 agcataaatg ggggattcac cagctttctg aggagaaagt ggaagtttaa gaaggaacga
3841 aacaaaggat acaagcacca tgctgaggat gctttgatca tcgctaacgc ggactttatc
3901 tttaaggaat ggaaaaagct ggataaggca aagaaagtga tggaaaacca gatgttcgag
3961 gagaagcagg cagagtcaat gcctgagatc gagacagagc aggaatacaa ggaaattttc
4021 atcacccctc atcagattaa acacataaag gacttcaaag actataaata ctctcatagg
4081 gtggacaaaa aacccaatcg cgagctcatt aatgacaccc tgtactcaac acggaaggat
4141 gataaaggta ataccttgat tgtgaataat cttaatggat tgtatgacaa agataacgac
4201 aagctcaaga agctgatcaa caagtctcca gagaagctcc ttatgtatca ccacgaccca
4261 cagacttatc agaaattgaa actgatcatg gagcaatacg gggatgagaa gaacccactc
4321 tacaaatatt atgaggaaac aggtaattac ctgaccaagt actccaagaa ggataacgga
4381 ccagtgatca aaaagataaa gtactatggc aacaaactta atgcgcattt ggacataact
4441 gacgattacc ccaattctcg aaacaaggtt gtgaagctct ccctgaagcc ttatagattt
4501 gacgtgtacc tggataatgg ggtttataaa ttcgtcaccg tgaaaaatct ggacgtgatc
4561 aaaaaggaga actattatga agtaaactca aagtgctatg aggaggcgaa gaagctgaag
4621 aagatctcca atcaggccga gttcatcgct tccttctata ataacgatct catcaagatc
4681 aatggagagc tttatcgcgt cattggtgtg aacaatgact tgctgaacag gatcgaagtc
4741 aatatgatag acattaccta ccgggagtat ctcgaaaaca tgaatgataa acggccgcct
4801 cgcatcatca agacaatcgc atctaaaact cagtcaataa aaaagtactc taccgatatc
4861 ctggggaatc tctatgaagt gaagtcaaag aagcacccac aaatcattaa aaaaggtgga
4921 tcccccaaga agaagaggaa agtctcgagc gactacaaag accatgacgg tgattataaa
4981 gatcatgaca tcgattacaa ggatgacgat gacaagtctg gtggttctgg aggatctggt
5041 ggttctacta atctgtcaga tattattgaa aaggagaccg gtaagcaact ggttatccag
5101 gaatccatcc tcatgctccc agaggaggtg gaagaagtca ttgggaacaa gccggaaagc
5161 gatatactcg tgcacaccgc ctacgacgag agcaccgacg agaatgtcat gcttctgact
5221 agcgacgccc ctgaatacaa gccttgggct ctggtcatac aggatagcaa cggtgagaac
5281 aagattaaga tgctctctgg tggttctgga ggatctggtg gttctactaa tctgtcagat
5341 attattgaaa aggagaccgg taagcaactg gttatccagg aatccatcct catgctccca
5401 gaggaggtgg aagaagtcat tgggaacaag ccggaaagcg atatactcgt gcacaccgcc
5461 tacgacgaga gcaccgacga gaatgtcatg cttctgacta gcgacgcccc tgaatacaag
5521 ccttgggctc tggtcataca ggatagcaac ggtgagaaca agattaagat gctctctggt
5581 ggttctccca agaagaagag gaaagtctaa ccggtcatca tcaccatcac cattgagttt
5641 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
5701 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
5761 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
5821 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
5881 ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga
5941 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
6001 cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct
6061 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
6121 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
6181 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
6241 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
6301 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
6361 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
6421 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
6481 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
6541 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
6601 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
6661 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
6721 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
6781 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
6841 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
6901 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
6961 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
7021 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
7081 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
7141 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
7201 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
7261 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
7321 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
7381 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
7441 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
7501 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
7561 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
7621 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
7681 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
7741 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
7801 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
7861 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
7921 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
7981 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
8041 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
8101 tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg tcgactctca
8161 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
8221 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
8281 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
8341 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
8401 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
8461 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
8521 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
8581 tggcagtaca tcaagtgtat c
//
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