| 质粒名称: | CRY2low-mCherry |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 104065 |
| 存储实验室: | Bianxiao Cui |
|
载体骨架基本信息 |
|
| 载体名称: | pmCherryN1 |
| 空载体大小: | 4000 bp |
| 完整质粒大小: | 6200 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: | CRY2low-mCherry/CRY2PHR/Cherry |
| 物种来源: | A. thaliana (mustard weed)芥花(芥菜杂草) |
| 插入大小: | 951 bp |
| 基因突变: | CRY2 aa 1-491 truncation, R489E A491D |
| 启动子: | CMV |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CMV forward |
|
文献引用方法 |
|
| 材料和方法部分: | CRY2low-mCherry was a gift from Bianxiao Cui (Addgene plasmid # 104065 ; http://n2t.net/addgene:104065 ; RRID:Addgene_104065) |
| 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
CRY2low-mCherry载体质粒基本信息
CRY2low-mCherry质粒图谱载体图谱和CRY2low-mCherry载体序列质粒序列多克隆位点信息
CRY2low-mCherry质粒载体简介
CRY2low-mCherry质粒序列载体序列
LOCUS CRY2low-mCherry 6167 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses fusion of CRY2low mutant (CRY2 aa1-491 R489E A491D) with
mCherry.
ACCESSION .
VERSION .
KEYWORDS CRY2low-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6167)
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 6167)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6167
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 73..376
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 377..580
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 531..551
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 2128..2137
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2134..2844
/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(2279..2297)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2547..2566
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2967..3088
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3004..3023)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3058..3077
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3095..3550)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3232..3253)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3444..3463
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3577..3681
/gene="bla"
/label=AmpR promoter
promoter 3683..4040
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3706..3726)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3891..4026
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3953..3972
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4075..4869
/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(4129..4148)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4739..4758
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5057..5076)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5101..5148
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5477..6065
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5966..5985
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
601 ccgctagcgc taccggactc agatctcgag gccaccatga agatggacaa aaagactata
661 gtttggttta gaagagacct aaggattgag gataatcctg cattagcagc agctgctcac
721 gaaggatctg tttttcctgt cttcatttgg tgtcctgaag aagaaggaca gttttatcct
781 ggaagagctt caagatggtg gatgaaacaa tcacttgctc acttatctca atccttgaag
841 gctcttggat ctgacctcac tttaatcaaa acccacaaca cgatttcagc gatcttggat
901 tgtatccgcg ttaccggtgc tacaaaagtc gtctttaacc acctctatga tcctgtttcg
961 ttagttcggg accataccgt aaaggagaag ctggtggaac gtgggatctc tgtgcaaagc
1021 tacaatggag atctattgta tgaaccgtgg gagatatact gcgaaaaggg caaacctttt
1081 acgagtttca attcttactg gaagaaatgc ttagatatgt cgattgaatc cgttatgctt
1141 cctcctcctt ggcggttgat gccaataact gcagcggctg aagcgatttg ggcgtgttcg
1201 attgaagaac tagggctgga gaatgaggcc gagaaaccga gcaatgcgtt gttaactaga
1261 gcttggtctc caggatggag caatgctgat aagttactaa atgagttcat cgagaagcag
1321 ttgatagatt atgcaaagaa cagcaagaaa gttgttggga attctacttc actactttct
1381 ccgtatctcc atttcgggga aataagcgtc agacacgttt tccagtgtgc ccggatgaaa
1441 caaattatat gggcaagaga taagaacagt gaaggagaag aaagtgcaga tctttttctt
1501 aggggaatcg gtttaagaga gtattctcgg tatatatgtt tcaacttccc gtttactcac
1561 gagcaatcgt tgttgagtca tcttcggttt ttcccttggg atgctgatgt tgataagttc
1621 aaggcctgga gacaaggcag gaccggttat ccgttggtgg atgccggaat gagagagctt
1681 tgggctaccg gatggatgca taacagaata agagtgattg tttcaagctt tgctgtgaag
1741 tttcttctcc ttccatggaa atggggaatg aagtatttct gggatacact tttggatgct
1801 gatttggaat gtgacatcct tggctggcag tatatctctg ggagtatccc cgatggccac
1861 gagcttgatc gcttggacaa tcccgcgtta caaggcgcca aatatgaccc agaaggtgag
1921 tacataaggc aatggcttcc cgagcttgcg agattgccaa ctgaatggat ccatcatcca
1981 tgggacgctc ctttaaccgt actcaaagct tctggtgtgg aactcggaac aaactatgcg
2041 aaacccattg tagacatcga cacagctcgt gagctactag ctaaagctat ttcaagaacc
2101 gaggaagacg cccgggatcc accggtcgcc accatggtga gcaagggcga ggaggataac
2161 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
2221 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
2281 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
2341 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
2401 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
2461 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
2521 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
2581 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
2641 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
2701 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
2761 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
2821 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
2881 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
2941 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
3001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
3061 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
3121 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
3181 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
3241 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
3301 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
3361 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
3421 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
3481 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
3541 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
3601 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
3661 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
3721 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
3781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
3841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
3901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
3961 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
4021 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
4081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
4141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
4201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
4261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
4321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
4381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
4441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
4501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
4561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
4621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
4681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
4741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
4801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
4861 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
4921 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
4981 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
5041 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
5101 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
5161 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
5221 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
5281 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
5341 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
5401 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
5461 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
5521 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
5581 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
5641 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
5701 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
5761 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
5821 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
5881 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
5941 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
6001 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
6061 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
6121 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtatt
//
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CRY2low-mCherry质粒载体应用举例
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