| 质粒名称: | CRY2-mCherry-Miro1TM |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 102247 |
| 存储实验室: | Bianxiao Cui |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4000 bp |
| 完整质粒大小: | 6400 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | CRY2PHR-mCherry-Miro1TM,CRY2,Miro1,RHOT1 |
| 物种来源: | H. sapiens (human), A. thaliana (mustard weed) |
| 插入大小: | 2400 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV forward |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | CRY2-mCherry-Miro1TM was a gift from Bianxiao Cui (Addgene plasmid # 102247 ; http://n2t.net/addgene:102247 ; RRID:Addgene_102247) |
| 参考文献部分: | Optogenetic control of molecular motors and organelle distributions in cells. Duan L, Che D, Zhang K, Ong Q, Guo S, Cui B. Chem Biol. 2015 May 21;22(5):671-82. doi: 10.1016/j.chembiol.2015.04.014. Epub 2015 May 9. 10.1016/j.chembiol.2015.04.014 PubMed 25963241 |
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 CRY2-mCherry-Miro1TM 6380 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses fusion of CRY2PHR with mCherry and transmembrane domain of
Miro1.
ACCESSION .
VERSION .
KEYWORDS CRY2-mCherry-Miro1TM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6380)
AUTHORS Duan L, Che D, Zhang K, Ong Q, Guo S, Cui B
TITLE Optogenetic control of molecular motors and organelle distributions
in cells.
JOURNAL Chem Biol. 2015 May 21;22(5):671-82. doi:
10.1016/j.chembiol.2015.04.014. Epub 2015 May 9.
PUBMED 25963241
REFERENCE 2 (bases 1 to 6380)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6380
/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"
CDS 625..2118
/codon_start=1
/product="N-terminal photolyase homology region of
Arabidopsis thaliana cryptochrome 2"
/label=CRY2PHR
/note="binds to CIB1 upon exposure to blue light (Kennedy
et al., 2010)"
/translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
regulatory 2137..2146
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2143..2850
/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(2288..2306)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2556..2575
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 2979..3044
/label=MCS
/note="multiple cloning site"
polyA_signal 3168..3289
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3205..3224)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3259..3278
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3296..3751)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3433..3454)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3645..3664
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3778..3882
/gene="bla"
/label=AmpR promoter
promoter 3884..4241
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3907..3927)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4092..4227
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4154..4173
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4276..5070
/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(4330..4349)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4940..4959
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5258..5277)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5302..5349
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5678..6266
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6167..6186
/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 atctcgaggc caccatgaag atggacaaaa agactatagt ttggtttaga
661 agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt
721 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca
781 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct
841 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt
901 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac
961 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat
1021 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat
1081 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg
1141 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta
1201 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca
1261 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat
1321 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat
1381 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg
1441 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt
1501 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg
1561 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga
1621 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga
1681 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt
1741 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt
1801 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc
1861 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa
1921 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct
1981 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta
2041 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag
2101 atcatgatcg gagcagcagc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
2161 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
2221 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
2281 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
2341 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
2401 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
2461 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
2521 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
2581 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
2641 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
2701 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
2761 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
2821 tccaccggcg gcatggacga gctgtacaag ccgcacgtga cacaagctga cctcaagagc
2881 tccacgtttt ggcttcgagc aagttttggt gctactgttt ttgcagtttt gggctttgct
2941 atgtacaaag cattattgaa acagcgatga tgtacaagtc cggactcaga tctcgagctc
3001 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac
3061 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
3121 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
3181 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
3241 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
3301 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
3361 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
3421 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
3481 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
3541 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
3601 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
3661 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
3721 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
3781 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
3841 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
3901 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
3961 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
4021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
4081 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
4141 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
4201 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
4261 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
4321 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
4381 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
4441 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
4501 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
4561 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
4621 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
4681 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
4741 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
4801 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
4861 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
4921 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
4981 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
5041 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
5101 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
5161 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
5221 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
5281 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
5341 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
5401 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
5461 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
5521 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
5581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
5641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
5701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
5761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
5821 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
5881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
5941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
6001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
6061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
6121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
6181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
6241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
6301 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
6361 accgtattac cgccatgcat
//
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