质粒名称: | pVenus-N1 (804) |
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出品公司: | Addgene |
目录编号: | 61854 |
存储实验室: | Jonathon Pines |
载体骨架基本信息 |
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载体名称: | pEYFP-N1 |
载体出品公司: | Clontech |
完整质粒大小: | 4733bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | Kanamycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | 无 |
标签/融合蛋白: | Venus (C terminal on backbone) |
文献引用方法 |
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材料和方法部分: |
pVenus-N1 (804) was a gift from Jonathon Pines (Addgene plasmid # 61854 ; http://n2t.net/addgene:61854 ; RRID:Addgene_61854)
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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
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载体序列
LOCUS pVenus-N1 (804) 4733 bp DNA circular SYN 25-AUG-2024 DEFINITION empty backbone for tagging proteins at the C-terminus with Venus. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4733) TITLE Pines Lab plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 4733) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.biofeng.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" FEATURES Location/Qualifiers source 1..4733 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 129..432 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 433..636 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 587..607 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 659..739 /label=MCS /note="multiple cloning site" regulatory 741..750 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 747..1466 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(792..813) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1053..1072) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1400..1421 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1589..1710 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1626..1645) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1680..1699 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1717..2172) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1854..1875) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2066..2085 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2199..2303 /gene="bla" /label=AmpR promoter promoter 2305..2662 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 2305..2496 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(2328..2348) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2513..2648 /label=SV40 ori /note="SV40 origin of replication" CDS 2697..3491 /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 2697..3491 /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(2751..2770) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3361..3380 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3679..3698) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3723..3770 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4099..4687 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4588..4607 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 61 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 121 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 181 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 241 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 301 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 361 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 421 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 481 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 541 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 601 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc 661 tagcgctacc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc 721 gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg 781 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 841 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg 901 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct 961 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 1021 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 1081 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 1141 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 1201 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca 1261 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 1321 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc 1381 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 1441 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac 1501 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 1561 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 1621 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 1681 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt 1741 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg 1801 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt 1861 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct 1921 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt 1981 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa 2041 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc 2101 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc 2161 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 2221 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 2281 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca 2341 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 2401 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 2461 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 2521 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 2581 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 2641 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga 2701 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 2761 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 2821 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag 2881 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 2941 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 3001 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3061 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 3121 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 3181 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg 3241 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 3301 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 3361 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 3421 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 3481 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 3541 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 3601 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 3661 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat 3721 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg 3781 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata 3841 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct 3901 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta 3961 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 4021 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 4081 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 4141 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 4201 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 4261 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 4321 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 4381 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 4441 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 4501 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 4561 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 4621 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 4681 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggc //
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