| 质粒名称: | pEGFP-Q74 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 40262 |
| 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: | 4700bp |
| 完整质粒大小: | 5019 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl2 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | HTT partial exon 1 Q74 / huntingtin |
| 物种来源: | H. sapiens (human) |
| 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 (未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 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-Q74 was a gift from David Rubinsztein (Addgene plasmid # 40262 ; http://n2t.net/addgene:40262 ; RRID:Addgene_40262)
|
| 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 |
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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 pEGFP-Q74 5019 bp ds-DNA circular SYN 03-MAR-2023
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pEGFP-Q74
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5019)
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 5019)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 3, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5019
/organism="synthetic DNA construct"
/mol_type="other DNA"
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
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 620..1336
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 620..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 1814..1935
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1851..1870)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1905..1924
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1942..2397)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2079..2100)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2291..2310
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2424..2528
/gene="bla"
/label=AmpR promoter
promoter 2530..2887
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2553..2573)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2738..2873
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2800..2819
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2922..3716
/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(2976..2995)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3586..3605
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3904..3923)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3948..3995
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4324..4912
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4813..4832
/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 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
1501 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
1561 gcagcagcag cagcagcagc agcagcagca gcagcagcaa cagccgccac cgccgccgcc
1621 gccgccgccg cctcctcagc ttcctcagcc gccgaattct gcagtcgacg gtaccgcggg
1681 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
1741 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
1801 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
1861 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
1921 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
1981 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
2041 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
2101 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
2161 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
2221 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
2281 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
2341 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
2401 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
2461 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
2521 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
2581 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
2641 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
2701 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
2761 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
2821 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
2881 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
2941 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
3001 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
3061 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
3121 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
3181 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
3241 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
3301 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
3361 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
3421 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
3481 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
3541 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
3601 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
3661 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
3721 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
3781 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
3841 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
3901 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
3961 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
4021 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
4081 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
4141 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
4201 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
4261 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
4321 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
4381 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
4441 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
4501 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
4561 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
4621 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
4681 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
4741 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
4801 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
4861 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
4921 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
4981 tgcgttatcc cctgattctg tggataaccg tattaccgc
//
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