| 质粒名称: | pCmv-BEACON1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 171697 |
| 存储实验室: | Jia Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 完整质粒大小: | 8076 bp |
| 载体类型: | Mammalian Expression, CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | BEACON1 / pCMV-SV40NLS-hAPOBEC3A_W104A/Y132D-XTEN-dLbCpf1(D832A/E1006A/D1125A)-SV40NLS-SGGS-UGI-SV40NLS-op |
| 物种来源: | H. sapiens (human); S. pyogenes, Bacteriophage PBS2 |
| 插入大小: | 4674 bp |
| 基因突变: | hAPOBEC3A_W104A/Y132D |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Unknown (unknown if destroyed) |
|
文献引用方法 |
|
| 材料和方法部分: |
pCMV-BEACON1 was a gift from Jia Chen (Addgene plasmid # 171697 ; http://n2t.net/addgene:171697 ; RRID:Addgene_171697)
|
| 参考文献部分: | Cas12a Base Editors Induce Efficient and Specific Editing with Low DNA Damage Response. Wang X, Ding C, Yu W, Wang Y, He S, Yang B, Xiong YC, Wei J, Li J, Liang J, Lu Z, Zhu W, Wu J, Zhou Z, Huang X, Liu Z, Yang L, Chen J. Cell Rep. 2020 Jun 2;31(9):107723. doi: 10.1016/j.celrep.2020.107723. 10.1016/j.celrep.2020.107723 PubMed 32492431 |
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 pCmv-BEACON1 8076 bp ds-DNA circular SYN 09-AUG-2021
DEFINITION Expresses BEACON1 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pCMV-BEACON1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8076)
AUTHORS Wang X, Ding C, Yu W, Wang Y, He S, Yang B, Xiong YC, Wei J, Li J,
Liang J, Lu Z, Zhu W, Wu J, Zhou Z, Huang X, Liu Z, Yang L, Chen J
TITLE Cas12a Base Editors Induce Efficient and Specific Editing with Low
DNA Damage Response.
JOURNAL Cell Rep. 2020 Jun 2;31(9):107723. doi:
10.1016/j.celrep.2020.107723.
PUBMED 32492431
REFERENCE 2 (bases 1 to 8076)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 9, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8076
/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 412..432
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4768..4788
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4801..5049
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5062..5082
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5091..5108
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5131..5148)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5137..5361
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5432..5448)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5432..5448)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5445..5467)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5456..5472
/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(5480..5510)
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind complement(5663..5680)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5834..6422)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5914..5933)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6593..7453)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLFRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7216..7235
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7454..7558)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7637..7656)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 7828..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 gcccaagaag
421 aagaggaaag tcatggaggc atctccagca agcggaccaa ggcacctgat ggacccccac
481 atcttcacct ctaactttaa caatggcatc ggcaggcaca agacatacct gtgctatgag
541 gtggagcgcc tggacaatgg caccagcgtg aagatggatc agcacagagg cttcctgcac
601 aaccaggcca agaatctgct gtgcggcttc tacggccggc acgcagagct gagatttctg
661 gacctggtgc ctagcctgca gctggatcca gcccagatct atagggtgac ctggttcatc
721 agctggtccc catgcttttc cgccggatgt gcaggagagg tgcgcgcctt cctgcaggag
781 aacacacacg tgcggctgag aatctttgcc gcccggatct acgacgatga tcctctgtac
841 aaggaggccc tgcagatgct gagagacgca ggagcccagg tgtccatcat gacctatgat
901 gagttcaagc actgctggga cacatttgtg gatcaccagg gctgtccctt tcagccttgg
961 gacggactgg atgagcactc ccaggccctg tctggcaggc tgagggccat cctgcagaac
1021 cagggcaata gcggcagcga gactcccggg acctcagagt ccgccacacc cgaaagtatg
1081 agcaagctgg agaagtttac aaactgctac tccctgtcta agaccctgag gttcaaggcc
1141 atccctgtgg gcaagaccca ggagaacatc gacaataagc ggctgctggt ggaggacgag
1201 aagagagccg aggattataa gggcgtgaag aagctgctgg atcgctacta tctgtccttt
1261 atcaacgacg tgctgcactc tatcaagctg aagaatctga acaattacat cagcctgttc
1321 cggaagaaga caagaaccga gaaggagaat aaggagctgg agaacctgga gatcaatctg
1381 cggaaggaga tcgccaaggc cttcaagggc aacgagggct acaagtccct gtttaagaag
1441 gatatcatcg agacaatcct gccagagttc ctggacgata aggacgagat cgccctggtg
1501 aacagcttca atggctttac cacagccttc accggcttct ttgataacag agagaatatg
1561 ttttccgagg aggccaagag cacatccatc gccttcaggt gtatcaacga gaatctgacc
1621 cgctacatct ctaatatgga catcttcgag aaggtggacg ccatctttga taagcacgag
1681 gtgcaggaga tcaaggagaa gatcctgaac agcgactatg atgtggagga tttctttgag
1741 ggcgagttct ttaactttgt gctgacacag gagggcatcg acgtgtacaa tgccatcatc
1801 ggcggcttcg tgaccgagag cggcgagaag atcaagggcc tgaacgagta catcaacctg
1861 tataatcaga agaccaagca gaagctgcct aagtttaagc cactgtataa gcaggtgctg
1921 agcgatcggg agtctctgag cttctacggc gagggctata catccgatga ggaggtgctg
1981 gaggtgttta gaaacaccct gaacaagaat tccgagatct tcagctccat caagaagctg
2041 gagaagctgt tcaagaattt tgacgagtac tctagcgccg gcatctttgt gaagaacggc
2101 cccgccatca gcacaatctc caaggatatc ttcggcgagt ggaacgtgat ccgggacaag
2161 tggaatgccg agtatgacga tatccacctg aagaagaagg ccgtggtgac cgagaagtac
2221 gaggacgatc ggagaaagtc cttcaagaag atcggctcct tttctctgga gcagctgcag
2281 gagtacgccg acgccgatct gtctgtggtg gagaagctga aggagatcat catccagaag
2341 gtggatgaga tctacaaggt gtatggctcc tctgagaagc tgttcgacgc cgattttgtg
2401 ctggagaaga gcctgaagaa gaacgacgcc gtggtggcca tcatgaagga cctgctggat
2461 tctgtgaaga gcttcgagaa ttacatcaag gccttctttg gcgagggcaa ggagacaaac
2521 agggacgagt ccttctatgg cgattttgtg ctggcctacg acatcctgct gaaggtggac
2581 cacatctacg atgccatccg caattatgtg acccagaagc cctactctaa ggataagttc
2641 aagctgtatt ttcagaaccc tcagttcatg ggcggctggg acaaggataa ggagacagac
2701 tatcgggcca ccatcctgag atacggctcc aagtactatc tggccatcat ggataagaag
2761 tacgccaagt gcctgcagaa gatcgacaag gacgatgtga acggcaatta cgagaagatc
2821 aactataagc tgctgcccgg ccctaataag atgctgccaa aggtgttctt ttccaagaag
2881 tggatggcct actataaccc ctctgaggac atccagaaga tctacaagaa tggcacattc
2941 aagaagggcg atatgtttaa cctgaatgac tgtcacaagc tgatcgactt ctttaaggat
3001 agcatctccc ggtatccaaa gtggagcaat gcctacgatt tcaacttttc cgagaccgag
3061 aagtataagg acatcgccgg cttttacaga gaggtggagg agcagggcta taaggtgagc
3121 ttcgagtctg ccagcaagaa ggaggtggat aagctggtgg aggagggcaa gctgtacatg
3181 ttccagatct ataacaagga cttttccgat aagtctcacg gcacacccaa tctgcacacc
3241 atgtacttca agctgctgtt tgacgagaac aatcacggcc agatcagact gtctggagga
3301 gcagagctgt tcatgaggag ggcaagcctg aagaaggagg agctggtggt gcacccagca
3361 aactccccta tcgccaacaa gaatccagat aatcccaaga agaccacaac cctgagctac
3421 gacgtgtata aggataagag gttttccgag gaccagtacg agctgcacat cccaatcgcc
3481 atcaataagt gccccaagaa catcttcaag atcaatacag aggtgcgcgt gctgctgaag
3541 cacgacgata acccctatgt gatcggcatc gccaggggcg agcgcaatct gctgtacatc
3601 gtggtggtgg acggcaaggg caacatcgtg gagcagtatt ccctgaacga gatcatcaac
3661 aacttcaacg gcatcaggat caagacagat taccactctc tgctggacaa gaaggagaag
3721 gagaggttcg aggcccgcca gaactggacc tccatcgaga atatcaagga gctaaaggcc
3781 ggctatatct ctcaggtggt gcacaagatc tgcgagctgg tggagaagta cgatgccgtg
3841 atcgccctgg ccgacctgaa ctctggcttt aagaatagcc gcgtgaaggt ggagaagcag
3901 gtgtatcaga agttcgagaa gatgctgatc gataagctga actacatggt ggacaagaag
3961 tctaatcctt gtgcaacagg aggcgccctg aagggatatc agatcaccaa taagttcgag
4021 agctttaagt ccatgtctac ccagaacggc ttcatctttt acatccctgc ctggctgaca
4081 tccaagatcg atccatctac cggctttgtg aatctgctga agacaaagta taccagcatc
4141 gccgattcca agaagttcat cagctccttt gacaggatca tgtacgtgcc cgaggaggat
4201 ctgttcgagt ttgccctgga ctataagaac ttctctcgca cagacgccga ttacatcaag
4261 aagtggaagc tgtacagcta tggcaaccgg atcagaatct tccggaatcc taagaagaac
4321 aacgtgttcg actgggagga ggtgtgcctg accagcgcct ataaggagct gttcaacaag
4381 tacggcatca attatcagca gggcgatatc agagccctgc tgtgcgagca gtccgacaag
4441 gccttctact ctagctttat ggccctgatg tctctgatgc tgcagatgag gaatagcatc
4501 acaggccgca ccgatgtggc ctttctgatc agccctgtga agaactccga cggcatcttc
4561 tacgatagcc ggaactatga ggcccaggag aatgccatcc tgccaaagaa cgccgacgcc
4621 aatggcgcct ataacatcgc cagaaaggtg ctgtgggcca tcggccagtt caagaaggcc
4681 gaggacgaga agctggataa ggtgaagatc gccatctcca acaaggagtg gctggagtac
4741 gcccagacct ctgtgaagca cggcagcccc aagaagaagc ggaaggtgtc tggtggttct
4801 accaacctgt ctgacatcat cgagaaggag acaggcaagc agctggtcat ccaggagagc
4861 atcctgatgc tgccagagga ggtggaggaa gtgatcggca acaagcccga gtccgatatc
4921 ctggtgcaca ccgcctacga cgagagcaca gatgagaatg tgatgctgct gacctccgac
4981 gcccccgagt ataagccttg ggccctggtc atccaggatt ctaacggcga gaataagatc
5041 aagatgctgt ctggtggttc tcccaagaag aagaggaaag tctaaccggt catcatcacc
5101 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
5161 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
5221 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
5281 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
5341 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt
5401 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
5461 atccgctcac aattccacac aacatacgag ccgaaagcat aaagtgtaaa gcctagggtg
5521 cctaatgagt aagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
5581 aaaacctgtc gtgccagctg cattaataaa tcggccaacg cgcggggaaa ggcggtttgc
5641 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
5701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
5761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
5821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
5881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
5941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
6001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
6061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
6121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
6181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
6241 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
6301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
6361 ctagtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
6421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
6481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
6541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
6601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
6661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
6721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
6781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
6841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
6901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
6961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
7021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
7081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
7141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
7201 cggcgtcaat acggaataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
7261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
7321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
7381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
7441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
7501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
7561 catttccccg aaaagtgcca cctgacgtcg acggatcggg agattgatct cccgatcccc
7621 tagggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
7681 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
7741 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
7801 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
7861 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
7921 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
7981 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
8041 ggtaaactgc ccacttggca gtacatcaag tgtatc
//
