| 质粒名称: | CRY2high-mCherry |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 104063 |
| 存储实验室: | Bianxiao Cui |
|
载体骨架基本信息 |
|
| 载体名称: | pmCherry-N1 |
| 空载体大小: | 4000 bp |
| 完整质粒大小: | 6188 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | CRY2high-mCherry/CRY2PHR/mCherry |
| 物种来源: | A. thaliana (mustard weed) |
| 基因突变: | CRY2PHR E490R |
| 启动子: | CMV |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CMV-F |
|
文献引用方法 |
|
| 材料和方法部分: | CRY2high-mCherry was a gift from Bianxiao Cui (Addgene plasmid # 104063 ; http://n2t.net/addgene:104063 ; RRID:Addgene_104063) |
| 参考文献部分: | Understanding CRY2 interactions for optical control of intracellular signaling. Duan L, Hope J, Ong Q, Lou HY, Kim N, McCarthy C, Acero V, Lin MZ, Cui B. Nat Commun. 2017 Sep 15;8(1):547. doi: 10.1038/s41467-017-00648-8. 10.1038/s41467-017-00648-8 [pii] PubMed 28916751 |
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 CRY2high-mCherry 6188 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses fusion of CRY2high mutant (CRY2PHR E490R) with mCherry.
ACCESSION .
VERSION .
KEYWORDS CRY2high-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6188)
AUTHORS Duan L, Hope J, Ong Q, Lou HY, Kim N, McCarthy C, Acero V, Lin MZ,
Cui B
TITLE Understanding CRY2 interactions for optical control of intracellular
signaling.
JOURNAL Nat Commun. 2017 Sep 15;8(1):547. doi: 10.1038/s41467-017-00648-8.
PUBMED 28916751
REFERENCE 2 (bases 1 to 6188)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6188
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 59..362
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 363..566
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 517..537
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 623..2116
/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
SGVELGTNYAKPIVDIDTARELLAKAISRTRRAQIMIGAA"
regulatory 2135..2144
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2141..2851
/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(2286..2304)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2554..2573
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2974..3095
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3011..3030)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3065..3084
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3102..3557)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3239..3260)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3451..3470
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3584..3688
/gene="bla"
/label=AmpR promoter
promoter 3690..4047
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3713..3733)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3898..4033
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3960..3979
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4082..4876
/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(4136..4155)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4746..4765
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5064..5083)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5108..5155
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5484..6072
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5973..5992
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
601 ggactcagat ctcgaggcca ccatgaagat ggacaaaaag actatagttt ggtttagaag
661 agacctaagg attgaggata atcctgcatt agcagcagct gctcacgaag gatctgtttt
721 tcctgtcttc atttggtgtc ctgaagaaga aggacagttt tatcctggaa gagcttcaag
781 atggtggatg aaacaatcac ttgctcactt atctcaatcc ttgaaggctc ttggatctga
841 cctcacttta atcaaaaccc acaacacgat ttcagcgatc ttggattgta tccgcgttac
901 cggtgctaca aaagtcgtct ttaaccacct ctatgatcct gtttcgttag ttcgggacca
961 taccgtaaag gagaagctgg tggaacgtgg gatctctgtg caaagctaca atggagatct
1021 attgtatgaa ccgtgggaga tatactgcga aaagggcaaa ccttttacga gtttcaattc
1081 ttactggaag aaatgcttag atatgtcgat tgaatccgtt atgcttcctc ctccttggcg
1141 gttgatgcca ataactgcag cggctgaagc gatttgggcg tgttcgattg aagaactagg
1201 gctggagaat gaggccgaga aaccgagcaa tgcgttgtta actagagctt ggtctccagg
1261 atggagcaat gctgataagt tactaaatga gttcatcgag aagcagttga tagattatgc
1321 aaagaacagc aagaaagttg ttgggaattc tacttcacta ctttctccgt atctccattt
1381 cggggaaata agcgtcagac acgttttcca gtgtgcccgg atgaaacaaa ttatatgggc
1441 aagagataag aacagtgaag gagaagaaag tgcagatctt tttcttaggg gaatcggttt
1501 aagagagtat tctcggtata tatgtttcaa cttcccgttt actcacgagc aatcgttgtt
1561 gagtcatctt cggtttttcc cttgggatgc tgatgttgat aagttcaagg cctggagaca
1621 aggcaggacc ggttatccgt tggtggatgc cggaatgaga gagctttggg ctaccggatg
1681 gatgcataac agaataagag tgattgtttc aagctttgct gtgaagtttc ttctccttcc
1741 atggaaatgg ggaatgaagt atttctggga tacacttttg gatgctgatt tggaatgtga
1801 catccttggc tggcagtata tctctgggag tatccccgat ggccacgagc ttgatcgctt
1861 ggacaatccc gcgttacaag gcgccaaata tgacccagaa ggtgagtaca taaggcaatg
1921 gcttcccgag cttgcgagat tgccaactga atggatccat catccatggg acgctccttt
1981 aaccgtactc aaagcttctg gtgtggaact cggaacaaac tatgcgaaac ccattgtaga
2041 catcgacaca gctcgtgagc tactagctaa agctatttca agaacccgta gagcacagat
2101 catgatcgga gcagcagccc gggatccacc ggtcgccacc atggtgagca agggcgagga
2161 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
2221 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
2281 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
2341 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
2401 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
2461 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
2521 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
2581 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
2641 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
2701 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
2761 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
2821 caccggcggc atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc
2881 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
2941 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
3001 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
3061 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
3121 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
3181 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
3241 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
3301 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
3361 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
3421 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
3481 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
3541 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
3601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
3661 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
3721 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
3781 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
3841 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
3901 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
3961 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
4021 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
4081 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
4141 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
4201 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
4261 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
4321 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
4381 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
4441 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
4501 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
4561 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
4621 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
4681 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
4741 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
4801 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
4861 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
4921 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
4981 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
5041 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
5101 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
5161 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
5221 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
5281 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
5341 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
5401 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
5461 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
5521 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
5581 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
5641 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
5701 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
5761 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
5821 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
5881 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
5941 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
6001 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
6061 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
6121 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
6181 atgcatta
//
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