| 质粒名称: | pmScarlet-i_C1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 85044 |
| 存储实验室: | Dorus Gadella |
|
载体骨架基本信息 |
|
| 载体名称: | pC1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4000 bp |
| 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: | mScarlet-i |
| 物种来源: | 合成的 |
| 插入大小: | 701 bp |
| 基因突变: | -- |
| 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: | AgeI(未破坏) |
| 3' 克隆位点: | BsrGI(未破坏) |
| 5' 测序引物 | CMV_fwd_primer |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pmScarlet-i_C1 was a gift from Dorus Gadella (Addgene plasmid # 85044 ; http://n2t.net/addgene:85044 ; RRID:Addgene_85044) |
| 参考文献部分: | mScarlet: a bright monomeric red fluorescent protein for cellular imaging. Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M, Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr. Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074. 10.1038/nmeth.4074 PubMed 27869816 |
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pmScarlet-i_C1载体 pmScarlet-i_C1质粒基本信息
pmScarlet-i_C1质粒图谱 pmScarlet-i_C1载体图谱和pmScarlet-i_C1载体序列 pmScarlet-i_C1质粒序列多克隆位点信息
pmScarlet-i_C1质粒 pmScarlet-i_C1载体简介
pmScarlet-i_C1质粒序列 pmScarlet-i_C1载体序列
LOCUS pmScarlet-i_C1 4710 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression of mScarlet-i.
ACCESSION .
VERSION .
KEYWORDS pmScarlet-i_C1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4710)
AUTHORS Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M,
Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr
TITLE mScarlet: a bright monomeric red fluorescent protein for cellular
imaging.
JOURNAL Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074.
PUBMED 27869816
REFERENCE 2 (bases 1 to 4710)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4710
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 119..422
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 423..626
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 577..597
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 665..674
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 671..1366
/codon_start=1
/product="bright monomeric red fluorescent protein evolved
from a synthetic template (Bindels et al., 2016)"
/label=mScarlet-I
/note="matures faster than mScarlet"
/translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
STGGMDELYK"
primer_bind complement(804..822)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1072..1091
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 1367..1432
/label=MCS
/note="multiple cloning site"
polyA_signal 1556..1677
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1593..1612)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1647..1666
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1684..2139)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1821..1842)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2033..2052
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2166..2270
/gene="bla"
/label=AmpR promoter
promoter 2272..2629
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2295..2315)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2480..2615
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2542..2561
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2664..3458
/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(2718..2737)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3328..3347
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3646..3665)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3690..3737
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4066..4654
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4555..4574
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
661 ggtcgccacc atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt
721 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
781 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
841 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc
901 cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt
961 gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg
1021 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat
1081 gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt
1141 gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga
1201 cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga
1261 ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg
1321 ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtccg gactcagatc
1381 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
1441 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
1501 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
1561 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
1621 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
1681 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
1741 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
1801 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
1861 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
1921 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
1981 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
2041 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
2101 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
2161 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
2221 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
2281 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
2341 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
2401 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
2461 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
2521 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
2581 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
2641 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
2701 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
2761 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
2821 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
2881 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
2941 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
3001 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
3061 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
3121 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
3181 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
3241 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
3301 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
3361 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
3421 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
3481 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
3541 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
3601 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
3661 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
3721 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
3781 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
3841 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
3901 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
3961 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
4021 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
4081 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
4141 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
4201 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
4261 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
4321 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
4381 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
4441 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
4501 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
4561 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
4621 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
4681 gttcctggcc ttttgctggc cttttgctca
//
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