| 质粒名称: | ORFtag_LambdaN-SD_Frame1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 40261 |
| 存储实验室: | David Rubinsztein |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 4989 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin, 50 μg/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: | HTT partial exon 1 Q23、huntingtin |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 951 bp |
| 基因突变: | expanded poly-Q |
| 启动子: | CMV |
| 标签/融合蛋白: | EGFP (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII (not destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 5’测序引物 | EGFP-C |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-Q23 was a gift from David Rubinsztein (Addgene plasmid # 40261 ; http://n2t.net/addgene:40261 ; RRID:Addgene_40261) |
| 参考文献部分: | A molecular investigation of true dominance in Huntington's disease. Narain Y, Wyttenbach A, Rankin J, Furlong RA, Rubinsztein DC. J Med Genet. 1999 Oct;36(10):739-46. 10.1136/jmg.36.10.739 PubMed 10528852 |
买家导航
pEGFP-Q23载体质粒基本信息
pEGFP-Q23质粒图谱载体图谱和pEGFP-Q23载体序列质粒序列多克隆位点信息
pEGFP-Q23质粒载体简介
pEGFP-Q23质粒序列载体序列
LOCUS pEGFP-Q23 4866 bp DNA circular SYN 14-APR-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4866)
AUTHORS Narain Y, Wyttenbach A, Rankin J, Furlong RA, Rubinsztein DC
TITLE A molecular investigation of true dominance in Huntington's disease.
JOURNAL J Med Genet. 1999 Oct;36(10):739-46.
PUBMED 10528852
REFERENCE 2 (bases 1 to 4866)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Med
Genet."; date: "1999-10"; volume: "36(10)"; pages: "739-46"
COMMENT Sequence Label: pEGFP-Q23
FEATURES Location/Qualifiers
source 1..4866
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 614..623
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS join(620..622,623..625,626..1336)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(620..622,623..625,626..1336)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(665..686)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(926..945)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1273..1294
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1661..1782
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1698..1717)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1752..1771
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1789..2244)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1926..1947)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2138..2157
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2271..2375
/gene="bla"
/label=AmpR promoter
promoter 2377..2734
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2377..2568
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2400..2420)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2585..2720
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2769..3563
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 2769..3563
/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(2823..2842)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3433..3452
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3751..3770)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3795..3842
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4171..4759
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4660..4679
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
1321 ggacgagctg tacaagtccg gactcagatc tatgaaggcc ttcgagtccc tcaagtcctt
1381 ccagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
1441 gcagcaacag ccgccaccgc cgccgccgcc gccgccgcct cctcagcttc ctcagccgcc
1501 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat
1561 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
1621 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
1681 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
1741 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
1801 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
1861 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
1921 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
1981 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
2041 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
2101 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
2161 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
2221 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
2281 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
2341 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
2401 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
2461 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
2521 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
2581 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
2641 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
2701 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
2761 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
2821 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
2881 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
2941 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
3001 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
3061 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
3121 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
3181 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
3241 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
3301 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
3361 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
3421 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
3481 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
3541 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
3601 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
3661 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
3721 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
3781 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
3841 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
3901 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
3961 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
4021 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
4081 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
4141 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
4201 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
4261 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
4321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
4381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
4441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
4501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
4561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
4621 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
4681 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
4741 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
4801 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
4861 taccgc
//
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