| 质粒名称: | pIRESneo-EGFP-alpha Tubulin |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 12298 |
| 存储实验室: | Patricia Wadsworth |
|
载体骨架基本信息 |
|
| 载体名称: | pIRESneo2 |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5300 bp |
| 完整质粒大小: | -- |
| 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: | EGFP-alpha Tubulin,EGFP-Tubulin,k-alpha-1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2100 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | EGFP（插入件上的N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI(未破坏) |
| 3' 克隆位点: | BamHI (未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pIRESneo-EGFP-alpha Tubulin was a gift from Patricia Wadsworth (Addgene plasmid # 12298 ; http://n2t.net/addgene:12298 ; RRID:Addgene_12298) |
| 参考文献部分: | Cell cycle-dependent changes in microtubule dynamics in living cells expressing green fluorescent protein-alpha tubulin. Rusan NM, Fagerstrom CJ, Yvon AM, Wadsworth P. Mol Biol Cell. 2001 Apr . 12(4):971-80. 10.1091/mbc.12.4.971 PubMed 11294900 |
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 pIRESneo-EGFP-alpha Tubulin 7372 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pIRESneo-EGFP-alpha Tubulin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7372) AUTHORS Rusan NM, Fagerstrom CJ, Yvon AM, Wadsworth P TITLE Cell cycle-dependent changes in microtubule dynamics in living cells expressing green fluorescent protein-alpha tubulin. JOURNAL Mol Biol Cell. 2001 Apr . 12(4):971-80. PUBMED 11294900 REFERENCE 2 (bases 1 to 7372) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7372 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 92..471 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 472..675 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 626..646 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 720..739 /label=T7 /note="T7 promoter, forward primer" promoter 720..738 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 800..809 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 806..1522 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 806..1522 /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(851..872) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1112..1131) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1459..1480 /label=EGFP-C /note="EGFP, forward primer" intron 2971..3200 /label=chimeric intron /note="chimera between introns from adenovirus and immunoglobulin heavy chain genes" primer_bind 3183..3202 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 3272..3845 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3439..3456) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3666..3685 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 3881..4684 /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3944..3963) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4554..4573 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4716..4733) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4722..4946 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 4993..5012 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5072..5088) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5072..5088) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5085..5107) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5096..5112 /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(5120..5150) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5165..5186 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5303..5320) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5474..6062) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5554..5573) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6233..7093) /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 6856..6875 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7094..7198) /gene="bla" /label=AmpR promoter primer_bind complement(7273..7292) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg 61 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag 121 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 181 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 241 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 301 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 361 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 421 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg 481 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc 541 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa 601 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc 661 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt 721 aatacgactc actataggga gacccaagct tggtaccgag ctcggatcga tatctgcggc 781 ctagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 841 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 901 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 961 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 1021 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 1081 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 1141 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 1201 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 1261 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 1321 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 1381 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 1441 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 1501 cggcatggac gagctgtaca agtccggact cagatctcga gtgcgtgagt gcatctccat 1561 ccacgttggc caggctggtg tccagattgg caatgcctgc tgggagctct actgcctgga 1621 acacggcatc cagcccgatg gccagatgcc aagtgacaag accattgggg gaggagatga 1681 ctccttcaac accttcttca gtgagacggg cgctggcaag cacgtgcccc gggctgtgtt 1741 tgtagacttg gaacccacag tcattgatga agttcgcact ggcacctacc gccagctctt 1801 ccaccctgag cagctcatca caggcaagga agatgctgcc aataactatg cccgagggca 1861 ctacaccatt ggcaaggaga tcattgacct tgtgttggac cgaattcgca agctggctga 1921 ccagtgcacc ggtcttcagg gcttcttggt tttccacagc tttggtgggg gaactggttc 1981 tgggttcacc tccctgctca tggaacgtct ctcagttgat tatggcaaga agtccaagct 2041 ggagttctcc atttacccag caccccaggt ttccacagct gtagttgagc cctacaactc 2101 catcctcacc acccacacca ccctggagca ctctgattgt gccttcatgg tagacaatga 2161 ggccatctat gacatctgtc gtagaaacct cgatatcgag cgcccaacct acactaacct 2221 taaccgcctt attagccaga ttgtgtcctc catcactgct tccctgagat ttgatggagc 2281 cctgaatgtt gacctgacag aattccagac caacctggtg ccctaccccc gcatccactt 2341 ccctctggcc acatatgccc ctgtcatctc tgctgagaaa gcctaccatg aacagctttc 2401 tgtagcagag atcaccaatg cttgctttga gccagccaac cagatggtga aatgtgaccc 2461 tcgccatggt aaatacatgg cttgctgcct gttgtaccgt ggtgacgtgg ttcccaaaga 2521 tgtcaatgct gccattgcca ccatcaaaac caagcgcagc atccagtttg tggattggtg 2581 ccccactggc ttcaaggttg gcatcaacta ccagcctccc actgtggtgc ctggtggaga 2641 cctggccaag gtacagagag ctgtgtgcat gctgagcaac accacagcca ttgctgaggc 2701 ctgggctcgc ctggaccaca agtttgacct gatgtatgcc aagcgtgcct ttgttcactg 2761 gtacgtgggt gaggggatgg aggaaggcga gttttcagag gcccgtgaag atatggctgc 2821 ccttgagaag gattatgagg aggttggtgt ggattctgtt gaaggagagg gtgaggaaga 2881 aggagaggaa tactaaggat ccgcggccgc atagataact gatccagtgt gctggaatta 2941 attcgctgtc tgcgagggcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga 3001 cttctgcgct aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg 3061 cggtgatgcc tttgagggtg gccgcgtcca tctggtcaga aaagacaatc tttttgttgt 3121 caagcttgag gtgtggcagg cttgagatct ggccatacac ttgagtgaca atgacatcca 3181 ctttgccttt ctctccacag gtgtccactc ccaggtccaa ctgcaggtcg agcatgcatc 3241 tagggcggcc aattccgccc ctctcccccc ccccctctcc ctcccccccc cctaacgtta 3301 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca 3361 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca 3421 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg 3481 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc 3541 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata 3601 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag 3661 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc 3721 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt 3781 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat 3841 gataagcttg ccacaacccg ggataattcc tgcagccaat atgggatcgg ccattgaaca 3901 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 3961 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 4021 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 4081 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 4141 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 4201 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 4261 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 4321 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 4381 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgatgatct 4441 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4501 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 4561 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4621 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4681 ctgaggggat caattctcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca 4741 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 4801 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 4861 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 4921 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgag 4981 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5041 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5101 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5161 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5221 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5281 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 5341 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5401 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5461 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5521 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5581 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5641 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 5701 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5761 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5821 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5881 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 5941 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6001 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 6061 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6121 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 6181 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 6241 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 6301 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 6361 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6421 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6481 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6541 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6601 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6661 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6721 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6781 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6841 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6901 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6961 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 7021 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 7081 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7141 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 7201 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 7261 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc 7321 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa ca //
