| 质粒名称: | pKA-142 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 79835 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pFC15K |
| 载体出品公司: | Promega |
| 空载体大小: | 2800 bp |
| 完整质粒大小: | 5000 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PpsR2-mVenus-CAAX |
| 物种来源: | Rhodopseudomonas palustris |
| 插入大小: | 2200 bp |
| 基因突变: | RpPpsR2半胱氨酸439变为丝氨酸 |
| GenBank编号 | KX063612 CAE26978.1 |
| 启动子: | CMVd1 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AsisI(未破坏) |
| 3’克隆位点: | NotI(未破坏) |
| 5’测序引物 | ggtaggcgtgtacggtgggag |
| 3’测序引物 | GCATCACAAATTTCACAAATAAAGC |
|
文献引用方法 |
|
| 材料和方法部分: | pKA-142 was a gift from Vladislav Verkhusha (Addgene plasmid # 79835 ; http://n2t.net/addgene:79835 ; RRID:Addgene_79835) |
| 参考文献部分: | A bacterial phytochrome-based optogenetic system controllable with near-infrared light. Kaberniuk AA, Shemetov AA, Verkhusha VV. Nat Methods. 2016 May 9. doi: 10.1038/nmeth.3864. 10.1038/nmeth.3864 PubMed 27159085 |
买家导航
pKA142载体基本信息
pKA142载体图谱质粒图谱和多克隆位点信息
pKA142载体简介
pKA142载体序列
LOCUS pKA-142 4987 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses PpsR2-mVenus-CAAX under CMVd1 promoter.
ACCESSION .
VERSION .
KEYWORDS pKA-142
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4987)
AUTHORS Kaberniuk AA, Shemetov AA, Verkhusha VV
TITLE A bacterial phytochrome-based optogenetic system controllable with
near-infrared light.
JOURNAL Nat Methods. 2016 May 9. doi: 10.1038/nmeth.3864.
PUBMED 27159085
REFERENCE 2 (bases 1 to 4987)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4987
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..100
/label=CMVd1 promoter
/note="truncated human cytomegalovirus (CMV) immediate
early promoter that yields slightly reduced expression
relative to the full-length promoter"
primer_bind 51..71
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 132..151
/label=T7
/note="T7 promoter, forward primer"
promoter 132..150
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 156..174
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 156..173
/label=SP6
/note="SP6 promoter, forward primer"
regulatory 1661..1670
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1667..2383
/codon_start=1
/product="Venus YFP with monomerizing A206K mutation (Nagai
et al., 2002; Kremers et al., 2006)"
/label=mVenus
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
ERVTAAGITLGMDELYK"
primer_bind complement(1712..1733)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1973..1992)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
polyA_signal 2461..2582
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2472..2491)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2526..2545
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS 2981..3775
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3035..3054)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3645..3664
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
rep_origin complement(3947..4535)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4027..4046)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
misc_feature 4651..4934
/label=cer region
/note="ColE1-derived recombination site that helps to
maintain plasmids as monomers"
ORIGIN
1 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
61 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
121 cactagaagc ttaatacgac tcactatagg gtacgattta ggtgacacta tagaataagg
181 agcgatcgcc atggctagcg cgtcaaagtc cgttcatgcc gacatcaccc ttctgctcga
241 tatggagggt gtgattcgcg aagccaccct gtctccgacg atggcggccg agagcgtgga
301 cggttggctg gggcgtcgct ggagcgacat cgccggcgcc gaaggcggcg acaaggttcg
361 ccgcatggtc gaagacgccc gccgcagcgg catctcggct ttccgccaga tcaatcagcc
421 tttcccgagc ggcgtcgaaa tcccgatcga attcaccacg atgctgctgg gcgaccgcac
481 cggcatgatc gcggtcggca agaacatgca ggcggtcacc gagctgcatt cccggctgat
541 cgctgcgcag caggcgatgg agcgcgacta ttggcggttg cgtgaattgg agactcgcta
601 ccgcctggtg ttcgacgctg ccgccgatgc ggtgatgatc gtctccgccg gcgacatgcg
661 catcgtcgaa gccaaccggg cggcggtgaa tgcgatcagc cgcgtcgagc gcggcaatga
721 cgaccttgcg gggcgtgatt tcctcgccga agtggcggct gccgatcgcg atgcggtgcg
781 cgacatgctg gcccaggtgc gtcagcgcgg caccgcactc agcgtcctcg ttcatctcgg
841 ccgttacgac cgcgcctgga tgctgcgcgg ttcgctgatg tcgtccgagc gtcgtcaggt
901 tttcctgctg cacttcaccc cggtgaccac gactcccgcg atcgacgacg tcgacgatga
961 tgccgtgctg cgcgggctga tcgatcgcat tcccgacggg ttcgtcgcac tggattcgga
1021 aggcgtcgtt cgtcacgcca accaggcgtt tctcgatctg gtccagatcg gctccaagcc
1081 tgcggcggtc ggacgatcgc tgggcgtctg gatgggtcgt ccgggcgccg atctgtccag
1141 cttgctgacg ctgctgcggc gctacaagac ggtgcggctg ttccaaacga cgatccgcgg
1201 cgagctcggc accgagactg aagtcgaggt ctcggccgtc gacggcgagg acgaccaata
1261 catcggcgtt ctgatgcgca atgtcgcgcg acgcctcgac gctgcggacg accacgatgc
1321 cttgcgtcag gcgctcggcc cgatcagcaa gcagctcggg cgatcctcgc tgcgcaagct
1381 ggtgaagaac gccgtgagca ttgtcgagca gcactacgtg aaggaagcgc tgttgcgatc
1441 caagggcaat cgcacggcaa ctgccgaact gctcggattg agccggcaga gcctttatgc
1501 aaaactcaac tcctacggct tcgacgacaa aggtgtcgtt gcttctgctg ccgacggtgc
1561 agagggcgcc tcagacgacg cagaggataa gcttagtgct ggtggtagtg ctggtggtag
1621 tgctggtggt agtgctggtg gtagtgctgg tggaccggtc gccaccatgg tgagcaaggg
1681 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
1741 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
1801 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
1861 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
1921 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
1981 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
2041 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
2101 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
2161 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
2221 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc taagctacca
2281 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggaacgcgt
2341 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggctagcg gtaaaaagaa
2401 gaaaaagaag tcaaagacaa agtgtgtaat tatgtaagcg gccgcttcga gcagacatga
2461 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
2521 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
2581 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
2641 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attctaatgg atcctctttg
2701 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc
2761 gtttctgcgg actggctttc tacgtgttcc gcttccttta gcagcccttg cgccctgagt
2821 gcttgcggca gcgtgagctt caaaagaatt gccagctggg gcgccctctg gtaaggttgg
2881 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
2941 atcaagatct gatcaagaga caggatgacg gtcgtttcgc atgcttgaac aagatggatt
3001 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
3061 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
3121 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
3181 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
3241 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
3301 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
3361 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgcactcg
3421 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
3481 agccgaactg ttcgccaggc tcaaggcgcg tatgccggat ggtgaggatc tcgtcgtgac
3541 tcatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
3601 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
3661 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
3721 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
3781 actctggggt tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
3841 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
3901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
3961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
4021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
4081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
4141 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
4201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
4261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
4321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
4381 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
4441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
4501 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
4561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
4621 aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc
4681 ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa
4741 tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat
4801 tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct
4861 gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
4921 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
4981 gagatct
//
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