| 质粒名称: | pMyosin-PSmOrange |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 31921 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pN1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | PSmOrange |
| 物种来源: | -- |
| 插入大小: | 711 bp |
| 基因突变: | S21T, Q63L, F100Y, L125M, K166R, P192S in PSmOrange (mutations are relative to mOrange but numbering is relative to EGFP) |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | Myosin（插入端N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pMyosin-PSmOrange was a gift from Vladislav Verkhusha (Addgene plasmid # 31921 ; http://n2t.net/addgene:31921 ; RRID:Addgene_31921) |
| 参考文献部分: | A photoswitchable orange-to-far-red fluorescent protein, PSmOrange. Subach OM, Patterson GH, Ting LM, Wang Y, Condeelis JS, Verkhusha VV. Nat Methods. 2011 Jul 31;8(9):771-7. doi: 10.1038/nmeth.1664. 10.1038/nmeth.1664 PubMed 21804536 |
买家导航
pMyosinPSmOrange载体基本信息
pMyosinPSmOrange载体图谱质粒图谱和多克隆位点信息
pMyosinPSmOrange载体简介
pMyosinPSmOrange载体序列
LOCUS pMyosin-PSmOrange 5210 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMyosin-PSmOrange
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5210)
AUTHORS Subach OM, Patterson GH, Ting LM, Wang Y, Condeelis JS, Verkhusha VV
TITLE A photoswitchable orange-to-far-red fluorescent protein, PSmOrange.
JOURNAL Nat Methods. 2011 Jul 31;8(9):771-7. doi: 10.1038/nmeth.1664.
PUBMED 21804536
REFERENCE 2 (bases 1 to 5210)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5210
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 1159..1168
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1165..1875
/codon_start=1
/product="orange to far-red photoswitchable variant of the
monomeric fluorescent protein mOrange (Subach et al.,
2011)"
/label=PSmOrange
/note="mammalian codon-optimized"
/translation="MVSKGEENNMAIIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEG
FQTAKLKVTKGGPLPFAWDILSPLFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIRMRLKLKDGGHYTSEVKTTYKAKKSVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind 1772..1795
/label=DsRed1-C
/note="DsRed1, forward primer"
polyA_signal 1998..2119
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2035..2054)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2089..2108
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2126..2581)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2263..2284)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2475..2494
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2608..2712
/gene="bla"
/label=AmpR promoter
promoter 2714..3071
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2737..2757)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2922..3057
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2984..3003
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3106..3900
/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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3160..3179)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3770..3789
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4088..4107)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4132..4179
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4508..5096
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4997..5016
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttatg tcgagcaaga gagccaaggc caagaccacc
661 aagaagaggc cccagagggc tacgtccaat gtcttcgcca tgtttgacca gtcccagatc
721 caggagttta aggaggcctt caacatgatt gatcagaacc gcgatggctt cattgataag
781 gaggacctgc acgacatgtt ggcctctctg gggaagaacc ccacagacga gtacctggag
841 ggcatgatga acgaggcgcc agggcccatc aacttcacca tgttcctcac aatgtttggg
901 gagaagctga acggcacaga ccccgaggat gtgatccgca atgcctttgc ctgctttgat
961 gaggaggcct caggcttcat ccacgaggac cacctgaggg agctgctcac caccatgggc
1021 gaccgattca cggatgagga ggtggacgag atgtaccgcg aggcacccat tgataagaag
1081 ggcaacttca actacgtgga gttcactcgc atcctcaaac acggcgccaa ggacaaggac
1141 gacctggatc caccggtcgc caccatggtg agcaagggcg aggagaataa tatggccatc
1201 atcaaggagt tcatgcgctt caaggtgcgc atggagggca ccgtgaacgg ccacgagttc
1261 gagatcgagg gcgagggcga gggccgcccc tacgagggct ttcagaccgc taagctgaag
1321 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccctct cttcacctac
1381 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttcaa gctgtccttc
1441 cccgagggct tcaagtggga gcgcgtgatg aactacgagg acggcggcgt ggtgaccgtg
1501 acccaggact cctcactgca ggacggcgag ttcatctaca aggtgaagat gcgcggcacc
1561 aacttcccct ccgacggccc cgtgatgcag aagaagacca tgggctggga ggcctcctcc
1621 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaggatgag gctgaagctg
1681 aaggacggcg gccactacac ctccgaggtc aagaccacct acaaggccaa gaagtccgtg
1741 cagctgcccg gcgcctacat cgtcggcatc aagctggaca tcacctccca caacgaggac
1801 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
1861 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
1921 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
1981 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
2041 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
2101 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
2161 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
2221 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
2281 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
2341 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
2401 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
2461 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
2521 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
2581 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
2641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
2701 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
2761 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
2821 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2881 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
2941 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
3001 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
3061 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
3121 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
3181 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
3241 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
3301 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
3361 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
3421 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
3481 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
3541 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
3601 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
3661 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
3721 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
3781 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
3841 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
3901 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
3961 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
4021 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
4081 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
4141 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
4201 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
4261 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
4321 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
4381 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
4441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
4501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
4561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
4621 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
4681 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
4741 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
4801 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
4861 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
4921 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
4981 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
5041 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
5101 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
5161 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
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