| 质粒名称: | DsRed2-ER-5 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 55836 |
| 存储实验室: | Michael Davidson |
|
载体骨架基本信息 |
|
| 载体名称: | DsRed2 |
| 载体出品公司: | -- |
| 空载体大小: | 4750 bp |
| 完整质粒大小: | 4757 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素，50μg/mL |
| 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: | ER |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 951 bp |
| 基因突变: | ER Signal Peptide |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | DsRed2 |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | DsRed2-ER-5 was a gift from Michael Davidson (Addgene plasmid # 55836 ; http://n2t.net/addgene:55836 ; RRID:Addgene_55836) |
| 参考文献部分: | The fluorescent protein palette: tools for cellular imaging. Day RN, Davidson MW. Chem Soc Rev. 2009 Oct;38(10):2887-921. doi: 10.1039/b901966a. Epub 2009 Aug 4. 10.1039/b901966a PubMed 19771335 |
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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 DsRed2-ER-5 4757 bp ds-DNA circular SYN 18-MAY-2023
DEFINITION Localization: Endoplasmic Reticulum, Excitation: 558, Emission: 583.
ACCESSION .
VERSION .
KEYWORDS DsRed2-ER-5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4757)
AUTHORS Day RN, Davidson MW
TITLE The fluorescent protein palette: tools for cellular imaging.
JOURNAL Chem Soc Rev. 2009 Oct;38(10):2887-921. doi: 10.1039/b901966a. Epub
2009 Aug 4.
PUBMED 19771335
REFERENCE 2 (bases 1 to 4757)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 18, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4757
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 30..134
/gene="bla"
/label=AmpR promoter
promoter 136..493
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(159..179)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 344..479
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 406..425
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 528..1322
/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(582..601)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1192..1211
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(1510..1529)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 1554..1601
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 1930..2518
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2419..2438
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 2693..2996
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2997..3200
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3151..3171
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 3295..3969
/codon_start=1
/product="improved tetrameric variant of DsRed fluorescent
protein"
/label=DsRed2
/note="mammalian codon-optimized"
/translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
LFL"
primer_bind complement(3475..3495)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 3887..3910
/label=DsRed1-C
/note="DsRed1, forward primer"
misc_feature 3997..4053
/label=MCS
/note="multiple cloning site"
polyA_signal 4177..4298
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4214..4233)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4268..4287
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(4305..3)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4442..4463)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4654..4673
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
61 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
121 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
241 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
301 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
361 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
421 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
481 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
541 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
601 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
661 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
721 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
781 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
841 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
901 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
961 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
1021 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
1081 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
1141 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
1201 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
1261 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
1321 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
1381 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
1441 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
1501 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
1561 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
1621 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
1681 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
1741 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
1801 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
1861 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
1921 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
1981 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
2041 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
2101 ccaccacttc aagaactctg tatcaccgcc tacatacctc gctctgctaa tcctgttacc
2161 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
2221 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
2281 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
2341 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
2401 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
2461 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
2521 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
2581 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
2641 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
2701 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
2761 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
2821 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
2881 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
2941 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
3001 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
3061 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
3121 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
3181 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcat gctgctatcc
3241 gtgccgttgc tgctcggcct cctcggcctg gccgtcgccg accggtcgca caccatggcc
3301 tcctccgaga acgtcatcac cgagttcatg cgcttcaagg tgcgcatgga gggcaccgtg
3361 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggccacaac
3421 accgtgaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
3481 ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac
3541 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
3601 ggcgtggcga ccgtgaccca ggactcctcc ctgcaggacg gctgcttcat ctacaaggtg
3661 aagttcatcg gcgtgaactt cccctccgac ggccccgtga tgcagaagaa gaccatgggc
3721 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagacccac
3781 aaggccctga agctgaagga cggcggccac tacctggtgg agttcaagtc catctacatg
3841 gccaagaagc ccgtgcagct gcccggctac tactacgtgg acgccaagct ggacatcacc
3901 tcccacaacg aggactacac catcgtggag cagtacgagc gcaccgaggg ccgccaccac
3961 ctgttcctga gatcgtacaa gaaggacgag ctgtaaagat ctcgagctca agcttcgaat
4021 tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc
4081 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
4141 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
4201 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
4261 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
4321 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
4381 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
4441 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
4501 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
4561 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
4621 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
4681 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
4741 tgcgccgcta cagggcg
//
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