质粒名称: | pcDNA-GFP |
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出品公司: | Addgene |
目录编号: | 160697 |
存储实验室: | Christine Mayr |
载体骨架基本信息 |
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载体名称: | pcDNA3.1 |
载体出品公司: | Clontech |
完整质粒大小: | 6188bp |
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: | 未知 |
插入基因信息 |
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插入基因名称: | GFP |
文献引用方法 |
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材料和方法部分: |
pcDNA-GFP was a gift from Christine Mayr (Addgene plasmid # 160697 ; http://n2t.net/addgene:160697 ; RRID:Addgene_160697)
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参考文献部分: | Gain of Additional BIRC3 Protein Functions through 3'-UTR-Mediated Protein Complex Formation. Lee SH, Mayr C. Mol Cell. 2019 May 16;74(4):701-712.e9. doi: 10.1016/j.molcel.2019.03.006. Epub 2019 Apr 1. 10.1016/j.molcel.2019.03.006 PubMed 30948266 |
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LOCUS pcDNA-GFP 6108 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses GFP in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6108) AUTHORS Lee SH, Mayr C TITLE Gain of Additional BIRC3 Protein Functions through 3'-UTR-Mediated Protein Complex Formation. JOURNAL Mol Cell. 2019 May 16;74(4):701-712.e9. doi: 10.1016/j.molcel.2019.03.006. Epub 2019 Apr 1. PUBMED 30948266 REFERENCE 2 (bases 1 to 6108) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6108 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 701..1420 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(746..767) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1007..1026) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1354..1375 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1465..1482) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1471..1695 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1741..2169 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1828..1847) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2038..2059 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2178..2198) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2183..2512 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2363..2498 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2425..2444 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2579..3373 /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(2633..2652) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3243..3262 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3547..3668 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3584..3603) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3638..3657 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3717..3733) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3717..3733) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3730..3752) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3741..3757 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3765..3795) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3810..3831 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3948..3965) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4119..4707) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4199..4218) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4878..5738) /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 5501..5520 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5739..5843) /gene="bla" /label=AmpR promoter primer_bind complement(5918..5937) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc atggtgagca agggcgagga 721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa 781 gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt 841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta 901 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc 961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta 1021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa 1081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa 1141 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa 1201 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac 1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc 1321 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc 1381 cgccgggatc actctcggca tggacgagct gtacaagtaa gcggccgctc gagtctagag 1441 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg 1501 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct 1561 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg 1621 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg 1681 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc 1741 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 1801 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 1861 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 1921 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 1981 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 2041 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 2101 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 2161 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 2281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 2341 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 2401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga 2461 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc 2521 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat 2581 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 2641 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 2701 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 2761 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 2821 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 2881 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 2941 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 3001 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 3061 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 3121 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 3181 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 3241 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 3301 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 3361 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg 3421 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 3481 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 3541 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 3601 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 3661 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca 3721 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 3781 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 3841 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 3901 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 3961 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 4021 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4081 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4141 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 4201 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 4261 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 4321 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 4381 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 4441 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 4501 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 4561 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 4621 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 4681 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 4741 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 4801 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 4861 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 4921 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 4981 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 5041 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 5101 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 5161 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 5221 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 5281 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 5341 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 5401 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 5461 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 5521 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 5581 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 5641 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 5701 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 5761 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 5821 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga 5881 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat 5941 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca 6001 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg 6061 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtt //
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