| 质粒名称: | CD63-pEGFP C2 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 62964 |
| 存储实验室: | Paul Luzio |
|
载体骨架基本信息 |
|
| 载体名称: | pN1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 5463 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | CD63 molecule |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 749 bp |
| 基因突变: | CD63 has 1 silent mutation compared to the coding sequence of NM_001267698.1 N129 AAT instead of AAC |
| GenBank编号 | NM_001267698.1 |
| 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: | KpnI(未破坏) |
| 3’克隆位点: | BamHI(未破坏) |
| 5’测序引物 | 5’ CCGACAACCACTACCTGAGC 3’ |
| 3’测序引物 | 5’ ACAAACCACAACTAGAATGCAG 3’ |
|
文献引用方法 |
|
| 材料和方法部分: | CD63-pEGFP C2 was a gift from Paul Luzio (Addgene plasmid # 62964 ; http://n2t.net/addgene:62964 ; RRID:Addgene_62964) |
买家导航
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 CD63-pEGFP C2 5463 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION full length human CD63 cloned into pEGFP C2.
ACCESSION .
VERSION .
KEYWORDS CD63-pEGFP C2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5463)
TITLE Luzio lab plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5463)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5463
/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"
CDS 1412..2128
/codon_start=1
/product="human CD63 tetraspanin"
/label=CD63
/note="enriched in intraluminal vesicles of late endosomes
(multivesicular bodies) and in exosomes"
/translation="MAVEGGMKCVKFLLYVLLLAFCACAVGLIAVGVGAQLVLSQTIIQ
GATPGSLLPVVIIAVGVFLFLVAFVGCCGACKENYCLMITFAIFLSLIMLVEVAAAIAG
YVFRDKVMSEFNNNFRQQMENYPKNNHTASILDRMQADFKCCGAANYTDWEKIPSMSKN
RVPDSCCINVTVGCGINFNEKAIHKEGCVEKIGGWLRKNVLVVAAAALGIAFVEVLGIV
FACCLVKSIRSGYEVM"
polyA_signal 2258..2379
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2295..2314)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2349..2368
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2386..2841)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2523..2544)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2735..2754
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2868..2972
/gene="bla"
/label=AmpR promoter
promoter 2974..3331
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2997..3017)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3182..3317
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3244..3263
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3366..4160
/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(3420..3439)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4030..4049
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4348..4367)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4392..4439
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4768..5356
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5257..5276
/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 gccggactca gatctcgagc tcaagcttcg aattctgcag
1381 tcgacggtac cagagcccag gcccggcagc catggcggtg gaaggaggaa tgaaatgtgt
1441 gaagttcttg ctctacgtcc tcctgctggc cttttgcgcc tgtgcagtgg gactgattgc
1501 cgtgggtgtc ggggcacagc ttgtcctgag tcagaccata atccaggggg ctacccctgg
1561 ctctctgttg ccagtggtca tcatcgcagt gggtgtcttc ctcttcctgg tggcttttgt
1621 gggctgctgc ggggcctgca aggagaacta ttgtcttatg atcacgtttg ccatctttct
1681 gtctcttatc atgttggtgg aggtggccgc agccattgct ggctatgtgt ttagagataa
1741 ggtgatgtca gagtttaata acaacttccg gcagcagatg gagaattacc cgaaaaataa
1801 ccacactgct tcgatcctgg acaggatgca ggcagatttt aagtgctgtg gggctgctaa
1861 ctacacagat tgggagaaaa tcccttccat gtcgaagaac cgagtccccg actcctgctg
1921 cattaatgtt actgtgggct gtgggattaa tttcaacgag aaggcgatcc ataaggaggg
1981 ctgtgtggag aagattgggg gctggctgag gaaaaatgtg ctggtggtag ctgcagcagc
2041 ccttggaatt gcttttgtcg aggttttggg aattgtcttt gcctgctgcc tcgtgaagag
2101 tatcagaagt ggctacgagg tgatgtaggg atccaccgga tctagataac tgatcataat
2161 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
2221 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
2281 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
2341 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
2401 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
2461 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
2521 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
2581 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
2641 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
2701 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
2761 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
2821 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
2881 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
2941 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
3001 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
3061 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
3121 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
3181 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
3241 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
3301 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
3361 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
3421 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
3481 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
3541 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
3601 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
3661 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
3721 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
3781 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
3841 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
3901 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
3961 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
4021 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
4081 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
4141 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
4201 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
4261 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
4321 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
4381 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
4441 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
4501 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
4561 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
4621 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
4681 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
4741 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
4801 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
4861 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
4921 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
4981 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
5041 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
5101 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
5161 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
5221 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
5281 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
5341 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
5401 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
5461 cgc
//
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