| 质粒名称: | pEGFP hsp70 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 15215 |
| 存储实验室: | Lois Greene |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C3 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4727 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: | hsp70 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | M11717 |
| 启动子 | -- |
| 标签/融合蛋白: | EGFP（主干上的N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII(未破坏) |
| 3' 克隆位点: | BamHI (未破坏) |
| 5' 测序引物 | EGFP-C primer |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP hsp70 was a gift from Lois Greene (Addgene plasmid # 15215 ; http://n2t.net/addgene:15215 ; RRID:Addgene_15215) |
| 参考文献部分: | Hsp70 dynamics in vivo: effect of heat shock and protein aggregation. Zeng XC, Bhasin S, Wu X, Lee JG, Maffi S, Nichols CJ, Lee KJ, Taylor JP, Greene LE, Eisenberg E. J Cell Sci. 2004 Oct 1. 117(Pt 21):4991-5000. 10.1242/jcs.01373 PubMed 15367583 |
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pEGFP hsp70载体 pEGFPhsp70质粒基本信息
pEGFP hsp70质粒图谱 pEGFPhsp70载体图谱和pEGFP hsp70载体序列 pEGFPhsp70质粒序列多克隆位点信息
pEGFP hsp70质粒 pEGFPhsp70载体简介
pEGFP hsp70质粒序列 pEGFPhsp70载体序列
LOCUS pEGFP hsp70 6888 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pEGFP hsp70
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6888)
AUTHORS Zeng XC, Bhasin S, Wu X, Lee JG, Maffi S, Nichols CJ, Lee KJ, Taylor
JP, Greene LE, Eisenberg E
TITLE Hsp70 dynamics in vivo: effect of heat shock and protein
aggregation.
JOURNAL J Cell Sci. 2004 Oct 1. 117(Pt 21):4991-5000.
PUBMED 15367583
REFERENCE 2 (bases 1 to 6888)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6888
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 54..357
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 358..561
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 512..532
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 600..609
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 606..1322
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(651..672)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(912..931)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1259..1280
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 3669..3790
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3706..3725)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3760..3779
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3797..4252)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3934..3955)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4146..4165
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4279..4383
/gene="bla"
/label=AmpR promoter
promoter 4385..4742
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4408..4428)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4593..4728
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4655..4674
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4777..5571
/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(4831..4850)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5441..5460
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5759..5778)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5803..5850
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6179..6767
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6668..6687
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
601 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
661 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
781 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
1021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
1081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
1141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
1201 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
1261 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
1321 agtactcaga tctcgagctc aagcttatgg ccaaagccgc ggcgatcggc atcgacctgg
1381 gcaccaccta ctcctgcgtg ggggtgttcc aacacggcaa ggtggagatc atcgccaacg
1441 accagggcaa ccgcaccacc cccagctacg tggccttcac ggacaccgag cggctcatcg
1501 gggatgcggc caagaaccag gtggcgctga acccgcagaa caccgtgttt gacgcgaagc
1561 ggctgatcgg ccgcaagttc ggcgacccgg tggtgcagtc ggacatgaag cactggcctt
1621 tccaggtgat caacgacgga gacaagccca aggtgcaggt gagctacaag ggggagacca
1681 aggcattcta ccccgaggag atctcgtcca tggtgctgac caagatgaag gagatcgccg
1741 aggcgtacct gggctacccg gtgaccaacg cggtgatcac cgtgccggcc tacttcaacg
1801 actcgcagcg ccaggccacc aaggatgcgg gtgtgatcgc ggggctcaac gtgctgcgga
1861 tcatcaacga gcccacggcc gccgccatcg cctacggcct ggacagaacg ggcaaggggg
1921 agcgcaacgt gctcatcttt gacctgggcg ggggcacctt cgacgtgtcc atcctgacga
1981 tcgacgacgg catcttcgag gtgaaggcca cggccgggga cacccacctg ggtggggagg
2041 actttgacaa caggctggtg aaccacttcg tggaggagtt caagagaaaa cacaagaagg
2101 acatcagcca gaacaagcga gccgtgaggc ggctgcgcac cgcctgcgag agggccaaga
2161 ggaccctgtc gtccagcacc caggccagcc tggagatcga ctccctgttt gagggcatcg
2221 acttctacac gtccatcacc agggcgaggt tcgaggagct gtgctccgac ctgttccgaa
2281 gcaccctgga gcccgtggag aaggctctgc gcgacgccaa gctggacaag gcccagattc
2341 acgacctggt cctggtcggg ggctccaccc gcatccccaa ggtgcagaag ctgctgcagg
2401 acttcttcaa cgggcgcgac ctgaacaaga gcatcaaccc cgacgaggct gtggcctacg
2461 gggcggcggt gcaggcggcc atcctgatgg gggacaagtc cgagaacgtg caggacctgc
2521 tgctgctgga cgtggctccc ctgtcgctgg ggctggagac ggccggaggc gtgatgactg
2581 ccctgatcaa gcgcaactcc accatcccca ccaagcagac gcagatcttc accacctact
2641 ccgacaacca acccggggtg ctgatccagg tgtacgaggg cgagagggcc atgacgaaag
2701 acaacaatct gttggggcgc ttcgagctga gcggcatccc tccggccccc aggggcgtgc
2761 cccagatcga ggtgaccttc gacatcgatg ccaacggcat cctgaacgtc acggccacgg
2821 acaagagcac cggcaaggcc aacaagatca ccatcaccaa cgacaagggc cgcctgagca
2881 aggaggagat cgagcgcatg gtgcaggagg cggagaagta caaagcggag gacgaggtgc
2941 agcgcgagag ggtgtcagcc aagaacgccc tggagtccta cgccttcaac atgaagagcg
3001 ccgtggagga tgaggggctc aagggcaaga tcagcgaggc cgacaagaag aaggtgctgg
3061 acaagtgtca agaggtcatc tcgtggctgg acgccaacac cttggccgag aaggacgagt
3121 ttgagcacaa gaggaaggag ctggagcagg tgtgtaaccc catcatcagc ggactgtacc
3181 agggtgccgg tggtcccggg cctgggggct tcggggctca gggtcccaag ggagggtctg
3241 ggtcaggccc caccattgag gaggtagatt aggggccttt ccaagattgc tgtttttgtt
3301 ttggagcttc aagactttgc atttcctagt atttctgttt gtcagttctc aatttcctgt
3361 gtttgcaatg ttgaaatttt ttggtgaagt cggatccacc ggatctagat aactgatcat
3421 aatcagccat accacatttg tagaggaagg gcgaattcca gcacactggc ggccgttact
3481 agtggatccg agctcggtac caagcttcga attctgcagt cgacggtacc gcgggcccgg
3541 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
3601 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
3661 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
3721 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
3781 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
3841 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
3901 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
3961 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
4021 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
4081 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
4141 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
4201 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
4261 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
4321 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
4381 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
4441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
4501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
4561 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
4621 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
4681 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
4741 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
4801 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
4861 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
4921 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
4981 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
5041 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
5101 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
5161 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
5221 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
5281 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
5341 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
5401 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
5461 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
5521 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
5581 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
5641 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
5701 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
5761 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
5821 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
5881 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
5941 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
6001 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
6061 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
6121 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
6181 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
6241 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
6301 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
6361 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
6421 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
6481 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
6541 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
6601 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
6661 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
6721 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
6781 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
6841 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttat
//
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