| 质粒名称: | pKA-141 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 79832 |
| 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: | 3000 bp |
| 完整质粒大小: | 5800 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: | BphP1-mCherry,RpBphP1 |
| 物种来源: | Rhodopseudomonas palustris |
| 插入大小: | 3000 bp |
| GenBank编号 | NCBI基因rpa1537 KX063613 |
| 启动子: | CMVd1 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NheI(未破坏) |
| 3’克隆位点: | XbaI (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-141 was a gift from Vladislav Verkhusha (Addgene plasmid # 79832 ; http://n2t.net/addgene:79832 ; RRID:Addgene_79832) |
| 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
pKA141载体基本信息
pKA141载体图谱质粒图谱和多克隆位点信息
pKA141载体简介
pKA141载体序列
LOCUS pKA-141 5805 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses RpBphP1-mCherry.
ACCESSION .
VERSION .
KEYWORDS pKA-141
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5805)
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 5805)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5805
/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"
CDS 2435..3145
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(2580..2598)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2848..2867
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(3224..3242)
/label=T7 Term
/note="T7 terminator, reverse primer"
polyA_signal 3279..3400
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3290..3309)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3344..3363
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS 3799..4593
/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(3853..3872)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4463..4482
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
rep_origin complement(4765..5353)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4845..4864)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
misc_feature 5469..5752
/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 tggcaggtca tgcctctggc agccccgcat tcgggaccgc
241 cgatctttcg aattgcgaac gtgaagagat ccacctcgcc ggctcgatcc agccgcatgg
301 cgcgcttctg gtcgtcagcg agccggatca tcgcatcatc caggccagcg ccaacgccgc
361 ggaatttctg aatctcggaa gcgtgctcgg cgttccgctc gccgagatcg acggcgatct
421 gttgatcaag atcctgccgc atctcgatcc caccgccgaa ggcatgccgg tcgcggtgcg
481 ctgccggatc ggcaatccct ccacggagta cgacggtctg atgcatcggc ctccggaagg
541 cgggctgatc atcgagctcg aacgtgccgg cccgccgatc gatctgtccg gcacgctggc
601 gccggcgctg gagcggatcc gcacggcggg ctcgctgcgc gcgctgtgcg atgacaccgc
661 gctgctgttt cagcagtgca ccggctacga ccgggtgatg gtgtatcgct tcgacgagca
721 gggccacggc gaagtgttct ccgagcgcca cgtgcccggg ctcgaatcct atttcggcaa
781 ccgctatccg tcgtcggaca ttccgcagat ggcgcggcgg ctgtacgagc ggcagcgcgt
841 ccgcgtgctg gtcgacgtca gctatcagcc ggtgccgctg gagccgcggc tgtcgccgct
901 gaccgggcgc gatctcgaca tgtcgggctg cttcctgcgc tcgatgtcgc cgatccatct
961 gcagtacctg aagaacatgg gcgtgcgcgc caccctggtg gtgtcgctgg tggtcggcgg
1021 caagctgtgg ggcctggttg cctgtcacca ttatctgccg cgcttcatcc atttcgagct
1081 gcgggcgatc tgcgaactgc tcgccgaagc gatcgcgacg cggatcaccg cgcttgagag
1141 cttcgcgcag agccagtcgg agctgttcgt gcagcggctc gaacagcgca tgatcgaagc
1201 gatcacccgt gaaggcgatt ggcgcgcagc gattttcgac accagccaat cgatcctgca
1261 gccgctgcac gccgacggtt gcgcgctggt gtacgaagac cagatcagga ccatcggtga
1321 cgtaccttcc acgcaggatg ttcgcgagat cgccgggtgg ctcgatcgcc agccacgtgc
1381 ggcggtgacc tcgaccgcgt cgctcggtct cgacgtgccg gagctcgcgc atctgacgcg
1441 gatggcgagc ggcgtggtcg cggcgccgat ttcggatcat cgcggcgagt ttctgatgtg
1501 gttccgcccc gagcgcgtcc acaccgttac ctggggcggc gatccgaaga agccgttcac
1561 gatgggcgat acaccggcgg atctgtcgcc gcggcgctcc ttcgccaaat ggcatcaggt
1621 tgtcgaaggc acgtccgatc cgtggacggc cgccgatctc gccgcggctc gcaccatcgg
1681 tcagaccgtc gccgacatcg tgctgcaatt ccgcgcggtg cggacactga tcgcccgcga
1741 acagtacgaa cagttttcgt cccaggtgca cgcttcgatg cagccggtgc tgatcaccga
1801 cgccgaaggc cgcatcctgc tgatgaacga ctcgttccgc gacatgttgc cggcggggtc
1861 gccatccgcc gtccatctcg acgatctcgc cgggttcttc gtcgaatcga acgatttcct
1921 gcgcaacgtc gccgaactga tcgatcacgg ccgcgggtgg cgcggcgaag ttctgctgcg
1981 cggcgcaggt aatcgcccgt tgccgctggc agtgcgcgcc gatccggtga cgcgcacgga
2041 ggaccagtcg ctcggcttcg tgctgatctt cagcgacgct accgatcgtc gcaccgcaga
2101 tgccgcacgc acgcgtttcc aggaaggcat tcttgccagc gcacgtcccg gcgtgcggct
2161 cgactccaag tccgacctct tgcacgagaa gctgctgtcc gcgctggtcg agaacgcgca
2221 gcttgccgca ttggaaatta cttacggcgt cgagaccgga cgcatcgccg agctgctcga
2281 aggcgttcgc cagtcgatgc tgcgcaccgc cgaagtgctc ggccatctgg tgcagcacgc
2341 ggcgcgcacg gccggcagcg acagctcgag caatggctcg cagaacaaga aggaattcga
2401 tagtgctggt agtgctggta gtgctggtac tagtatggtg agcaagggcg aggaggataa
2461 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
2521 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
2581 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
2641 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
2701 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
2761 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
2821 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
2881 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
2941 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
3001 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
3061 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
3121 cggcatggac gagctgtaca agtaatctag agtcgacctg caggcatgca agctgatccg
3181 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
3241 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
3301 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
3361 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
3421 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
3481 caaatgtggt aaaatcgaat tctaatggat cctctttgcg cttgcgtttt cccttgtcca
3541 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
3601 cgtgttccgc ttcctttagc agcccttgcg ccctgagtgc ttgcggcagc gtgagcttca
3661 aaagaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg
3721 atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca
3781 ggatgacggt cgtttcgcat gcttgaacaa gatggattgc acgcaggttc tccggccgct
3841 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
3901 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
3961 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
4021 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
4081 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
4141 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
4201 caccaagcga aacatcgcat cgagcgagca cgcactcgga tggaagccgg tcttgtcgat
4261 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
4321 aaggcgcgta tgccggatgg tgaggatctc gtcgtgactc atggcgatgc ctgcttgccg
4381 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
4441 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
4501 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
4561 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
4621 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
4681 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
4741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
4801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
4861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
4921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
4981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
5041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
5101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
5161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
5221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
5281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
5341 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
5401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
5461 tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc
5521 atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg
5581 cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt
5641 tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta
5701 caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc
5761 gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatct
//
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