| 质粒名称: | pCI-neo.mCherry-NEDD4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 38316 |
| 存储实验室: | Quan Lu |
|
载体骨架基本信息 |
|
| 载体名称: | pCI-neo |
| 载体出品公司: | Promega |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5472 bp |
| 完整质粒大小: | 8855 bp |
| 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: | Nedd4 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 3411 bp |
| 基因突变: | -- |
| GenBank编号 | NM_006154.2 |
| 启动子 | CMV |
| 标签/融合蛋白: | mCherry（N端子插入） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI（克隆过程中销毁） |
| 3' 克隆位点: | NotI（克隆过程中销毁） |
| 5' 测序引物 | T7 promoter |
| 3' 测序引物 | T3 promoter |
|
文献引用方法 |
|
| 材料和方法部分: | pCI-neo.mCherry-NEDD4 was a gift from Quan Lu (Addgene plasmid # 38316 ; http://n2t.net/addgene:38316 ; RRID:Addgene_38316) |
| 参考文献部分: | Arrestin domain-containing protein 3 recruits the NEDD4 E3 ligase to mediate ubiquitination of the beta2-adrenergic receptor. Nabhan JF, Pan H, Lu Q. EMBO Rep. 2010 Aug;11(8):605-11. Epub 2010 Jun 18. 10.1038/embor.2010.80 PubMed 20559325 |
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载体基本信息
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载体简介
载体序列
LOCUS pCI-neo.mCherry-NEDD4 8855 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCI-neo.mCherry-NEDD4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8855) AUTHORS Nabhan JF, Pan H, Lu Q TITLE Arrestin domain-containing protein 3 recruits the NEDD4 E3 ligase to mediate ubiquitination of the beta2-adrenergic receptor. JOURNAL EMBO Rep. 2010 Aug;11(8):605-11. Epub 2010 Jun 18. PUBMED 20559325 REFERENCE 2 (bases 1 to 8855) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8855 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(10..27) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 45..166 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(56..75) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 110..129 /label=SV40pA-R /note="SV40 polyA, reverse primer" rep_origin 352..807 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(439..458) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 649..670 /label=F1ori-F /note="F1 origin, forward primer" promoter 924..1281 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(947..967) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1132..1267 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1194..1213 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 1332..2126 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1386..1405) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1996..2015 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 2190..2238 /note="synthetic polyadenylation signal" primer_bind complement(2279..2298) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2398..2420 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2458..2476) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2544..2648 /gene="bla" /label=AmpR promoter CDS 2649..3509 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2867..2886) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3680..4268 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4169..4188 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 4479..4858 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4859..5070 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5021..5041 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5067..5091 /label=LNCX /note="Human CMV promoter, forward primer" intron 5231..5363 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 5346..5365 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 5408..5426 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 5443..6150 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(5588..5606) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 5856..5875 /label=mCherry-F /note="mCherry, forward primer" ORIGIN 1 ggccgcttcc ctttagtgag ggttaatgct tcgagcagac atgataagat acattgatga 61 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga 121 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg 181 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa 241 cctctacaaa tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa tagcgaagag 301 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct 361 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 421 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 481 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 541 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 601 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 661 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 721 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 781 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg 841 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa 901 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag 961 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 1021 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 1081 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 1141 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 1201 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 1261 ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagg 1321 ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 1381 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 1441 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 1501 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 1561 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 1621 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 1681 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 1741 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 1801 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 1861 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 1921 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 1981 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 2041 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 2101 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 2161 gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt 2221 gtgttggttt tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc actctcagta 2281 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg 2341 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg 2401 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc 2461 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag 2521 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 2581 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 2641 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 2701 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 2761 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 2821 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 2881 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 2941 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 3001 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 3061 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 3121 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 3181 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 3241 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 3301 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 3361 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 3421 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 3481 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 3541 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 3601 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 3661 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 3721 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 3781 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 3841 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 3901 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 3961 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 4021 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 4081 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 4141 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 4201 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 4261 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 4321 ctcacatggc tcgacagatc ttcaatattg gccattagcc atattattca ttggttatat 4381 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta 4441 catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat tgactagtta 4501 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac 4561 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc 4621 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt 4681 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc 4741 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac 4801 cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt 4861 gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc 4921 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt 4981 tccaaaatgt cgtaacaact gcgatcgccc gccccgttga cgcaaatggg cggtaggcgt 5041 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cactagaagc 5101 tttattgcgg tagtttatca cagttaaatt gctaacgcag tcagtgcttc tgacacaaca 5161 gtctcgaact taagctgcag tgactctctt aaggtagcct tgcagaagtt ggtcgtgagg 5221 cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg 5281 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat 5341 ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct cttaaggcta 5401 gagtacttaa tacgactcac tataggctag cctcgagaat tcatggtgag caagggcgag 5461 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc 5521 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc 5581 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg 5641 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac 5701 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac 5761 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag 5821 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg 5881 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc 5941 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac 6001 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc 6061 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac 6121 tccaccggcg gcatggacga gctgtacaag atggcaactt gcgcggtgga ggtgttcggg 6181 ctcctggagg acgaggaaaa ttcacgaatt gtgagagtaa gagttatagc cggaataggc 6241 cttgccaaga aggatatatt gggagctagt gatccttacg tgagagtgac gttatatgac 6301 ccaatgaatg gagttcttac aagtgtgcaa acaaaaacca ttaaaaagag tttgaatcca 6361 aagtggaatg aagaaatatt attcagagtt catcctcagc agcaccggct tctttttgaa 6421 gtgtttgacg aaaaccgatt gacaagagat gatttcctag gtcaagtgga tgttccactt 6481 tatccattac cgacagaaaa tccaagattg gagagaccat atacatttaa ggattttgtt 6541 cttcatccaa gaagtcacaa atcaagagtt aaaggttatc tgagactaaa aatgacttat 6601 ttacctaaaa ccagtggctc agaagatgat aatgcagaac aggctgagga attagagcct 6661 ggctgggttg ttttggacca accagatgct gcttgccatt tgcagcaaca acaagaacct 6721 tctcctctac ctccagggtg ggaagagagg caggatatcc ttggaaggac ctattatgta 6781 aaccatgaat ctagaagaac acagtggaaa agaccaaccc ctcaggacaa cctaacagat 6841 gctgagaatg gcaacattca actgcaagca caacgtgcat ttaccaccag gcggcagata 6901 tccgaggaaa cagaaagtgt tgacaaccga gagtcttccg agaactggga aattataaga 6961 gaagatgaag ccaccatgta tagcaaccag gccttcccat cacctccacc gtcaagtaac 7021 ttggatgttc caactcatct tgcagaagaa ttgaatgcca gactcaccat ttttggaaat 7081 tcagccgtga gccagccagc atcgagctca aatcattcca gcagaagagg cagcttacaa 7141 gcctatactt ttgaggaaca acctacactt cctgtgcttt tgcctacttc atctggatta 7201 ccaccaggtt gggaagaaaa acaagatgaa agaggaagat catattatgt agatcacaat 7261 tccagaacga ctacttggac aaagcccact gtacaggcca cagtggagac cagtcagctg 7321 acctcaagcc agagttctgc aggccctcaa tcacaagcct ccaccagtga ttcaggccag 7381 caggtgaccc agccatctga aattgagcaa ggattccttc ctaaaggctg ggaagtccgg 7441 catgcaccaa atgggaggcc tttctttatt gaccacaaca ctaaaaccac cacctgggaa 7501 gatccaagat tgaaaattcc agcccatctg agaggaaaga catcacttga tacttccaat 7561 gatctagggc ctttacctcc aggatgggaa gagagaactc acacagatgg aagaatcttc 7621 tacataaatc acaatataaa aagaacacaa tgggaagatc ctcggttgga gaatgtagca 7681 ataactggac cagcagtgcc ctactccagg gattacaaaa gaaagtatga gttcttccga 7741 agaaagttga agaagcagaa tgacattcca aacaaatttg aaatgaaact tcgccgagca 7801 actgttcttg aagactctta ccggagaatt atgggtgtca agagagcaga cttcctgaag 7861 gctcgactgt ggattgagtt tgatggtgaa aagggattgg attatggagg agttgccaga 7921 gaatggttct tcctgatctc aaaggaaatg tttaaccctt attatgggtt gtttgaatat 7981 tctgctacgg acaattatac cctacagata aatccaaact ctggattgtg taacgaagat 8041 cacctctctt acttcaagtt tattggtcgg gtagctggaa tggcagttta tcatggcaaa 8101 ctgttggatg gttttttcat ccgcccattt tacaagatga tgcttcacaa accaataacc 8161 cttcatgata tggaatctgt ggatagtgaa tattacaatt ccctaagatg gattcttgaa 8221 aatgacccaa cagaattgga cctcaggttt atcatagatg aagaactttt tggacagaca 8281 catcaacatg agctgaaaaa tggtggatca gaaatagttg tcaccaataa gaacaaaaag 8341 gaatatattt atcttgtaat acaatggcga tttgtaaacc gaatccagaa gcaaatggct 8401 gcttttaaag agggattctt tgaactaata ccacaggatc tcatcaaaat ttttgatgaa 8461 aatgaactag agcttcttat gtgtggactg ggagatgttg atgtgaatga ctggagggaa 8521 catacaaagt ataaaaatgg ctacagtgca aatcatcagg ttatacagtg gttttggaag 8581 gctgttttaa tgatggattc agaaaaaaga ataagattac ttcagtttgt cactggcaca 8641 tctcgggtgc ctatgaatgg atttgctgaa ctatacggtt caaatggacc acagtcattt 8701 acagttgaac agtggggtac tcctgaaaag ctgccaagag ctcatacctg ttttaatcgc 8761 ctggacttgc caccttatga atcatttgaa gaattatggg ataaacttca gatggcaatt 8821 gaaaacaccc agggctttga tggagttgat taggc //
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