| 质粒名称: | EGFP-CAAX |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 86056 |
| 存储实验室: | Lei Lu |
|
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: | 4731bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | Homo sapiens Harvey rat sarcoma |
| 物种来源: | H. sapiens (human) |
| 基因突变: | the region from 825-857 of HRAS as a farnesylation motif |
| GenBank编号 | NM_001318054.1 |
| 启动子: | CMV |
| 标签/融合蛋白: | GFP (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI (not destroyed) |
| 3’克隆位点: | BamHI (not destroyed) |
| 5’测序引物 | TTTAGTGAACCGTCAGATC |
|
文献引用方法 |
|
| 材料和方法部分: | EGFP-CAAX was a gift from Lei Lu (Addgene plasmid # 86056 ; http://n2t.net/addgene:86056 ; RRID:Addgene_86056) |
| 参考文献部分: | A ternary complex comprising transportin1, Rab8 and the ciliary targeting signal directs proteins to ciliary membranes. Madugula V, Lu L. J Cell Sci. 2016 Oct 15;129(20):3922-3934. Epub 2016 Sep 15. 10.1242/jcs.194019 PubMed 27633000 |
买家导航
EGFP-CAAX载体质粒基本信息
EGFP-CAAX质粒图谱载体图谱和EGFP-CAAX载体序列质粒序列多克隆位点信息
EGFP-CAAX质粒载体简介
EGFP-CAAX质粒序列载体序列
LOCUS EGFP-CAAX 4740 bp DNA circular SYN 14-APR-2025
DEFINITION A mammalian expression plasmid encoding farnesylation motif of HRAS
with N-terminus EGFP..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4740)
AUTHORS Madugula V, Lu L
TITLE A ternary complex comprising transportin1, Rab8 and the ciliary
targeting signal directs proteins to ciliary membranes.
JOURNAL J Cell Sci. 2016 Oct 15;129(20):3922-3934. Epub 2016 Sep 15.
PUBMED 27633000
REFERENCE 2 (bases 1 to 4740)
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 Cell
Sci."; date: "2016-10-15"; volume: "129(20)"; pages: "3922-3934.
Epub 2016 Sep 15"
COMMENT Sequence Label: EGFP-CAAX
FEATURES Location/Qualifiers
source 1..4740
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 9..597
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 498..517
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 772..1075
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1076..1279
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1230..1250
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 1318..1327
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS join(1324..1326,1327..1329,1330..2040)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(1324..1326,1327..1329,1330..2040)
/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(1369..1390)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1630..1649)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1977..1998
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2239..2360
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2276..2295)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2330..2349
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2367..2822)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2504..2525)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2716..2735
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2849..2953
/gene="bla"
/label=AmpR promoter
promoter 2955..3312
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2955..3146
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2978..2998)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3163..3298
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3347..4141
/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 3347..4141
/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(3401..3420)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4011..4030
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4329..4348)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4373..4420
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
721 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
781 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
841 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
901 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
961 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
1021 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
1081 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
1141 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
1201 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
1261 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc
1321 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
1381 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
1441 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
1501 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
1561 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
1621 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
1681 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
1741 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
1801 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
1861 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
1921 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
1981 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
2041 tccggactca gatctcgagc tcaagcttcg aattcaggct gcatgagctg caagtgtgtg
2101 ctctcctgag gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
2161 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
2221 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
2281 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
2341 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
2401 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
2461 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
2521 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
2581 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
2641 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
2701 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
2761 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
2821 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
2881 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
2941 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
3001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
3061 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
3121 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
3181 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
3241 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
3301 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
3361 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
3421 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
3481 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
3541 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
3601 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
3661 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
3721 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
3781 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
3841 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
3901 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
3961 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
4021 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
4081 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
4141 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
4201 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
4261 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
4321 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
4381 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
4441 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
4501 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
4561 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
4621 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
4681 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
//
