| 质粒名称: | CRY2olig-mCherry |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 60032 |
| 存储实验室: | Chandra Tucker |
|
载体骨架基本信息 |
|
| 载体名称: | pmCherryN1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4000 bp |
| 完整质粒大小: | 6187 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | CRY2PHR E490G,CRY2 |
| 物种来源: | A. thaliana (mustard weed) |
| 插入大小: | 1495 bp |
| 基因突变: | E490G |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XhoI (未破坏) |
| 3’克隆位点: | XmaI(未破坏) |
| 5’测序引物 | CMV |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | CRY2olig-mCherry was a gift from Chandra Tucker (Addgene plasmid # 60032 ; http://n2t.net/addgene:60032 ; RRID:Addgene_60032) |
| 参考文献部分: | An optimized optogenetic clustering tool for probing protein interaction and function. Taslimi A, Vrana JD, Chen D, Borinskaya S, Mayer BJ, Kennedy MJ, Tucker CL. Nat Commun. 2014 Sep 18;5:4925. doi: 10.1038/ncomms5925. 10.1038/ncomms5925 PubMed 25233328 |
买家导航
CRY2oligmCherry载体基本信息
CRY2oligmCherry载体图谱质粒图谱和多克隆位点信息
CRY2oligmCherry载体简介
CRY2oligmCherry载体序列
LOCUS CRY2olig-mCherry 6188 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses a fusion of CRY2PHR E490G with mCherry.
ACCESSION .
VERSION .
KEYWORDS CRY2olig-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6188)
AUTHORS Taslimi A, Vrana JD, Chen D, Borinskaya S, Mayer BJ, Kennedy MJ,
Tucker CL
TITLE An optimized optogenetic clustering tool for probing protein
interaction and function.
JOURNAL Nat Commun. 2014 Sep 18;5:4925. doi: 10.1038/ncomms5925.
PUBMED 25233328
REFERENCE 2 (bases 1 to 6188)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6188
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 87..444
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(110..130)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 295..430
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 357..376
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 479..1273
/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(533..552)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1143..1162
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(1461..1480)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 1505..1552
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 1881..2469
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2370..2389
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 2644..2947
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2948..3151
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3102..3122
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 3208..4701
/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
SGVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAA"
regulatory 4720..4729
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4726..5436
/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(4871..4889)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 5139..5158
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 5559..5680
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5596..5615)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5650..5669
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(5687..6142)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5824..5845)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6036..6055
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 6169..85
/gene="bla"
/label=AmpR promoter
ORIGIN
1 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
61 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
121 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
181 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
241 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
301 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
361 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
421 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
481 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
541 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
601 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
661 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
721 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
781 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
841 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
901 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
961 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
1021 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
1081 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
1141 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
1201 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
1261 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
1321 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
1381 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
1441 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
1501 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
1561 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
1621 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
1681 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
1741 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
1801 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
1861 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
1921 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
1981 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
2041 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
2101 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
2161 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
2221 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
2281 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
2341 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
2401 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
2461 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
2521 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
2581 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
2641 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
2701 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
2761 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
2821 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
2881 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
2941 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
3001 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
3061 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
3121 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
3181 ctaccggact cagatctcga ggccaccatg aagatggaca aaaagactat agtttggttt
3241 agaagagacc taaggattga ggataatcct gcattagcag cagctgctca cgaaggatct
3301 gtttttcctg tcttcatttg gtgtcctgaa gaagaaggac agttttatcc tggaagagct
3361 tcaagatggt ggatgaaaca atcacttgct cacttatctc aatccttgaa ggctcttgga
3421 tctgacctca ctttaatcaa aacccacaac acgatttcag cgatcttgga ttgtatccgc
3481 gttaccggtg ctacaaaagt cgtctttaac cacctctatg atcctgtttc gttagttcgg
3541 gaccataccg taaaggagaa gctggtggaa cgtgggatct ctgtgcaaag ctacaatgga
3601 gatctattgt atgaaccgtg ggagatatac tgcgaaaagg gcaaaccttt tacgagtttc
3661 aattcttact ggaagaaatg cttagatatg tcgattgaat ccgttatgct tcctcctcct
3721 tggcggttga tgccaataac tgcagcggct gaagcgattt gggcgtgttc gattgaagaa
3781 ctagggctgg agaatgaggc cgagaaaccg agcaatgcgt tgttaactag agcttggtct
3841 ccaggatgga gcaatgctga taagttacta aatgagttca tcgagaagca gttgatagat
3901 tatgcaaaga acagcaagaa agttgttggg aattctactt cactactttc tccgtatctc
3961 catttcgggg aaataagcgt cagacacgtt ttccagtgtg cccggatgaa acaaattata
4021 tgggcaagag ataagaacag tgaaggagaa gaaagtgcag atctttttct taggggaatc
4081 ggtttaagag agtattctcg gtatatatgt ttcaacttcc cgtttactca cgagcaatcg
4141 ttgttgagtc atcttcggtt tttcccttgg gatgctgatg ttgataagtt caaggcctgg
4201 agacaaggca ggaccggtta tccgttggtg gatgccggaa tgagagagct ttgggctacc
4261 ggatggatgc ataacagaat aagagtgatt gtttcaagct ttgctgtgaa gtttcttctc
4321 cttccatgga aatggggaat gaagtatttc tgggatacac ttttggatgc tgatttggaa
4381 tgtgacatcc ttggctggca gtatatctct gggagtatcc ccgatggcca cgagcttgat
4441 cgcttggaca atcccgcgtt acaaggcgcc aaatatgacc cagaaggtga gtacataagg
4501 caatggcttc ccgagcttgc gagattgcca actgaatgga tccatcatcc atgggacgct
4561 cctttaaccg tactcaaagc ttctggtgtg gaactcggaa caaactatgc gaaacccatt
4621 gtagacatcg acacagctcg tgagctacta gctaaagcta tttcaagaac ccgtggagca
4681 cagatcatga tcggagcagc agcccgggat ccaccggtcg ccaccatggt gagcaagggc
4741 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
4801 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
4861 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
4921 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
4981 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
5041 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
5101 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
5161 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
5221 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
5281 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
5341 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
5401 cactccaccg gcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
5461 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
5521 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
5581 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
5641 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
5701 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
5761 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
5821 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
5881 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
5941 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
6001 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
6061 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
6121 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
6181 atttgttt
//
CRY2oligmCherry载体图谱质粒图谱pdf版和相关资料下载
CRY2oligmCherry载体质粒应用举例
- 上一篇：FUW-tetO-hOKMS
- 下一篇：pDIRECT_22C
