| 质粒名称: | pC0091 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 181955 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pC0037 |
| 空载体大小: | 6957 bp |
| 完整质粒大小: | 6309 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | 7xFOP / Mutant 7xTOP binding site |
| 物种来源: | Synthetic |
| 插入大小: | 149 bp |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | ctagcaaaataggctgtccc |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pC0091 was a gift from Feng Zhang (Addgene plasmid # 181955 ; http://n2t.net/addgene:181955 ; RRID:Addgene_181955) |
| 参考文献部分: | A cytosine deaminase for programmable single-base RNA editing. Abudayyeh OO, Gootenberg JS, Franklin B, Koob J, Kellner MJ, Ladha A, Joung J, Kirchgatterer P, Cox DBT, Zhang F. Science. 2019 Jul 11. pii: science.aax7063. doi: 10.1126/science.aax7063. 10.1126/science.aax7063 PubMed 31296651 |
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 pC0091 6309 bp ds-DNA circular SYN 14-FEB-2022
DEFINITION Beta catenin reporter control M51 Super 8x (mutated TCF/LEF binding
sites) FOPFlash with Gluc/Cluc.
ACCESSION .
VERSION .
KEYWORDS pC0091
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6309)
AUTHORS Abudayyeh OO, Gootenberg JS, Franklin B, Koob J, Kellner MJ, Ladha
A, Joung J, Kirchgatterer P, Cox DBT, Zhang F
TITLE A cytosine deaminase for programmable single-base RNA editing.
JOURNAL Science. 2019 Jul 11. pii: science.aax7063. doi:
10.1126/science.aax7063.
PUBMED 31296651
REFERENCE 2 (bases 1 to 6309)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 14, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6309
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 50..69
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(288..392)
/gene="bla"
/label=AmpR promoter
rep_origin 419..874
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(506..525)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 716..737
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 1027..1075
/note="synthetic polyadenylation signal"
CDS complement(1076..2737)
/codon_start=1
/product="secreted Cypridina luciferase"
/label=CLuc
/note="contains 30 codon substitutions for efficient
translation in mammalian cells"
/translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
primer_bind complement(2762..2781)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2763..2781)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(2826..3029)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(2855..2875)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 3030..3409
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
polyA_signal 3436..3484
/note="synthetic polyadenylation signal"
misc_feature 3498..3589
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 3538..3557
/label=RVprimer3
/note="pGL3 vector, forward primer"
promoter 3746..3777
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 3833..3842
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3839..4396
/codon_start=1
/product="secreted Gaussia luciferase"
/label=hGLuc
/note="human codon-optimized"
/translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
RCATFASKIQGQVDKIKGAGGD"
polyA_signal 4437..4558
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4448..4467)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4502..4521
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(4806..4823)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4977..5565)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5057..5076)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5736..287)
/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"
ORIGIN
1 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
61 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
121 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
181 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
241 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
301 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
361 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
421 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
481 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
541 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
601 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
661 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
721 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
781 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
841 gcgaatttta acaaaatatt aacgcttaca atttgccatt cgccattcag gctgcgcaac
901 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa gctaccatga
961 taagtaagta atattaaggt acgggaggta cttggagcgg ccgcagctat gaccatgatt
1021 acgccacaca caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattttatt
1081 tgcattcatc tggtacttct agggtgtctc catgctttat gtagcattct ttcttgaact
1141 cccatgcgtg gtcacagaaa ccctgttgtc ccctcaggca atactcgtac atgcatcgtt
1201 cgacacggtc aggcttgtgg cacacgttac atttctgatc aagatcactt tgaccggcga
1261 agagactatt gcagagtcgt tcagcttcag gtttctgttc ttcggtgcat cccggtgggt
1321 tgggggtcag atcacaggct ccttcagcat caaaagaatc atcactgaaa tcctggttgt
1381 agttaccgca aataccgcaa gtcttaccat cgaatggatc tctaatatgt acgacgagca
1441 gttgcttgaa gttgaacttg accaccagag cttcaggtag gatggctgta gtcagtatgt
1501 caccatcttg ccagtagatg gaagtgttct gagagctgta cgggacggac acggcttctc
1561 ctccaaccag aatctgtttt ccatcaacaa tgagttctac tacagtcgat tgtttcctga
1621 ttcgtacttt ctctacttca gtgtaagagt caacattcct gtgtgaaacc ttcacatccc
1681 aagtgttcca gaaacagtcg gcggccataa gaatctcctt gcagggaccc tggaattggt
1741 atcttgcttt gtcaaatgta tcgtagaaag tatgtccaga caaaacgcag gttcctctag
1801 tttcgggagc agcgcacgtc tccctgtagt caagcagcac gtgtgaggct cgctcgtctc
1861 cacccataca gcgggcgaag gcgcaggaga tggtgtagta gtagaagttg atggggttgc
1921 ggcagctgtc cttgtacggc tccagaagac ctttgcagta tgcaacgtca tcaggatttc
1981 catagagtgg acaaccgtca aactcctggt tgatcttagg ctgaatagcg agttgttctg
2041 gatctgaagt gaagtctgta tcttccatca ttttaagatc tccacagagg ccagagatca
2101 ttcctttgtt tgctgtgtct ggggcgattc ttacagatct tcctccgagg atgtcgatca
2161 cgatcagttt gaagaactca atgacggtga tgttgaagcc tggcatctca acaacagcga
2221 tggtgacctc gccgatggtg tacgggttgg cgatgatagg gtcagctcca ccgtttacag
2281 tgatgggatt ctcagtagct tgagcgatgt caatgatgtc tccagccact tccagtcttg
2341 tcttggtcag cacagccccc ttggttccat ccaggttctc cagggtgatg gacaccttcc
2401 agtcgccgcc cttggttcct tgacccaaca cgtatgtacc aggttcctgg aactggaatc
2461 tctttccata gaatgttcta aaccatcctg cggcctctac tctgcattca attacatact
2521 gacacattcg acaacatgtt tttcctggtt tattttcaca cagtccatct gatagtatgt
2581 ctctcgtgca ggtgccacag ctgctatcaa tacattctcc ttctttagct tcacaggaag
2641 ttggaactgt gtttggtgga tcaggttcgt aaggacagtc ctggcaatga acagtggcgc
2701 agtagactaa tgcaacggca agaattaagg tcttcatggt ggcggtacca agcttgggtc
2761 tccctatagt gagtcgtatt aatttcgata agccagtaag cagtgggttc tctagttagc
2821 cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
2881 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
2941 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
3001 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
3061 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
3121 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
3181 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
3241 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
3301 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
3361 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat
3421 ctggcccgta catcgaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
3481 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
3541 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgataggta
3601 ccttacgcga ggccaaaggg ggtaaggcca aagggggtaa ggccaaaggg ggtaaggcca
3661 aaggcgcgag gccaaagggg gtaaggccaa agggggtaag gccaaagggg gtaaggccaa
3721 aggcccgggc tcgagatcta gactctagag ggtatataat ggaagctcga attccagctt
3781 ggcattccgg tactgttggt aaaaagcttg gcattccggt actgttggta aagccaccat
3841 gggagtcaaa gttctgtttg ccctgatctg catcgctgtg gccgaggcca agcccaccga
3901 gaacaacgaa gacttcaaca tcgtggccgt ggccagcaac ttcgcgacca cggatctcga
3961 tgctgaccgc gggaagttgc ccggcaagaa gctgccgctg gaggtgctca aagagatgga
4021 agccaatgcc cggaaagctg gctgcaccag gggctgtctg atctgcctgt cccacatcaa
4081 gtgcacgccc aagatgaaga agttcatccc aggacgctgc cacacctacg aaggcgacaa
4141 agagtccgca cagggcggca taggcgaggc gatcgtcgac attcctgaga ttcctgggtt
4201 caaggacttg gagcccatgg agcagttcat cgcacaggtc gatctgtgtg tggactgcac
4261 aactggctgc ctcaaagggc ttgccaacgt gcagtgttct gacctgctca agaagtggct
4321 gccgcaacgc tgtgcgacct ttgccagcaa gatccagggc caggtggaca agatcaaggg
4381 ggccggtggt gactaattct agagtcgggg cggccggccg cttcgagcag acatgataag
4441 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
4501 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
4561 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
4621 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct
4681 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
4741 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc
4801 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
4861 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
4921 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
4981 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
5041 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
5101 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
5161 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
5221 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
5281 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
5341 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
5401 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
5461 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
5521 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
5581 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
5641 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
5701 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
5761 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
5821 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
5881 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
5941 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
6001 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
6061 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
6121 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
6181 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
6241 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
6301 tcattctga
//
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