| 质粒名称: | pCI-neo.html' target='_blank'>pCIneoEGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 46949 |
| 存储实验室: | John Schiller |
|
载体骨架基本信息 |
|
| 载体名称: | pCIneo |
| 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: | 6220 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物的表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
| 物种来源: | 维多利亚多管发光水母 |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 未知 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | NA |
| 3' 测序引物 | NA |
|
文献引用方法 |
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| 材料和方法部分: | pCIneoEGFP was a gift from John Schiller (Addgene plasmid # 46949 ; http://n2t.net/addgene:46949 ; RRID:Addgene_46949) |
| 参考文献部分: | Production of papillomavirus-based gene transfer vectors. Buck CB, Thompson CD. Curr Protoc Cell Biol. 2007 Dec;Chapter 26:Unit 26.1. doi: 10.1002/0471143030.cb2601s37. 10.1002/0471143030.cb2601s37 PubMed 18228512 |
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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 pCI-neo.html' target='_blank'>pCIneoEGFP 6220 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION GFP in Promega pCI-neo.html' target='_blank'>pCIneo (uses CMV promoter). Superior to pfwB for some purposes. ACCESSION . VERSION . KEYWORDS pCIneoEGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6220) AUTHORS Buck CB, Thompson CD TITLE Production of papillomavirus-based gene transfer vectors. JOURNAL Curr Protoc Cell Biol. 2007 Dec;Chapter 26:Unit 26.1. doi: 10.1002/0471143030.cb2601s37. PUBMED 18228512 REFERENCE 2 (bases 1 to 6220) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6220 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 138..517 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 518..729 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 680..700 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 726..750 /label=LNCX /note="Human CMV promoter, forward primer" intron 890..1022 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 1005..1024 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 1067..1085 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 1091..1143 /label=MCS /note="multiple cloning site" regulatory 1151..1160 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1157..1876 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1202..1223) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1463..1482) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1810..1831 /label=EGFP-C /note="EGFP, forward primer" promoter complement(1889..1906) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 1924..2045 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1935..1954) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1989..2008 /label=SV40pA-R /note="SV40 polyA, reverse primer" rep_origin 2231..2686 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2318..2337) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2528..2549 /label=F1ori-F /note="F1 origin, forward primer" promoter 2803..3160 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2826..2846) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3011..3146 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3073..3092 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3211..4005 /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(3265..3284) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3875..3894 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4069..4117 /note="synthetic polyadenylation signal" primer_bind complement(4158..4177) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4277..4299 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4337..4355) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4423..4527 /gene="bla" /label=AmpR promoter CDS 4528..5388 /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(4746..4765) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5559..6147 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6048..6067 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta 61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg 181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg 661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata 721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac 781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt 841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa 901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact 961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac 1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact 1081 ataggctagc ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga 1141 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 1201 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 1261 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 1321 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 1381 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 1441 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 1501 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 1561 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 1621 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 1681 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 1741 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 1801 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 1861 cgagctgtac aagtaaagcg gccgcttccc tttagtgagg gttaatgctt cgagcagaca 1921 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct 1981 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac 2041 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg 2101 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat ccgataagga tcgatccggg 2161 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa 2221 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2281 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2341 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2401 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2461 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2521 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2581 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 2641 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt 2701 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg 2761 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca 2821 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 2881 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 2941 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 3001 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 3061 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 3121 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac 3181 aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt 3241 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 3301 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 3361 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg 3421 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 3481 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 3541 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 3601 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 3661 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 3721 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 3781 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 3841 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 3901 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 3961 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt 4021 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct 4081 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc 4141 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac 4201 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga 4261 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa 4321 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata 4381 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt 4441 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg 4501 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt 4561 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta 4621 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc 4681 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa 4741 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc 4801 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt 4861 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact 4921 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac 4981 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata 5041 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta 5101 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg 5161 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat 5221 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt 5281 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga 5341 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa 5401 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 5461 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 5521 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 5581 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 5641 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 5701 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 5761 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 5821 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 5881 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 5941 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 6001 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 6061 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 6121 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 6181 gccttttgct ggccttttgc tcacatggct cgacagatct //
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