质粒名称: | pcDNA3-myc-CUL3 |
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出品公司: | 爱迪基因 |
目录编号: | 19893 |
存储实验室: | Yue Xiong |
载体骨架基本信息 |
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载体名称: | pcDNA3-myc |
载体出品公司: | Invitrogen |
载体抗性: | 氨苄霉素 |
空载体大小: | 5512 bp |
完整质粒大小: | 7816 bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | G418 |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | cullin 3 |
物种来源: | 人源 |
插入大小: | 2304 bp |
基因又名: | CUL3 |
GenBank编号 | NM_003590 |
标签/融合蛋白: | 3xmyc (N terminal on backbone) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | KpnI(未破坏) |
3' 克隆位点: | XhoI(未破坏) |
5' 测序引物 | T7 |
3' 测序引物 | SP6 |
文献引用方法 |
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材料和方法部分: | pcDNA3-myc-CUL3 was a gift from Yue Xiong (爱迪基因 plasmid # 19893) |
参考文献部分: | ROC1, a homolog of APC11, represents a family of cullin partners with an associated ubiquitin ligase activity. Ohta T, Michel JJ, Schottelius AJ, Xiong Y. Mol Cell. 1999 Apr . 3(4):535-41. 10.1016/S1097-2765(00)80482-7 PubMed 10230407 |
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载体基本信息
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载体序列
LOCUS pcDNA3-myc-CUL3 8175 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-myc-CUL3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8175) AUTHORS Ohta T, Michel JJ, Schottelius AJ, Xiong Y TITLE ROC1, a homolog of APC11, represents a family of cullin partners with an associated ubiquitin ligase activity. JOURNAL Mol Cell. 1999 Apr . 3(4):535-41. PUBMED 10230407 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pcDNA3-myc-CUL3| FEATURES Location/Qualifiers source 1..8175 /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 663..692 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 699..728 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 735..764 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" promoter complement(3494..3512) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3495..3512) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3532..3549) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3538..3762 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3808..4236 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3895..3914) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4105..4126 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4245..4265) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4250..4579 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4430..4565 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4492..4511 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4646..5440 /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(4700..4719) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5310..5329 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5614..5735 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5651..5670) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5705..5724 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5784..5800) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5784..5800) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5797..5819) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5808..5824 /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(5832..5862) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5877..5898 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6015..6032) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6186..6774) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6266..6285) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6945..7805) /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 7568..7587 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7806..7910) /gene="bla" /label=AmpR promoter primer_bind complement(7985..8004) /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 acccaagcta 661 tggagcaaaa gctcatctca gaggaagatc tggaattaga gcaaaagctc atctcagagg 721 aagatctgga attagagcaa aagctcatct cagaggaaga tctggaattc aggatcctga 781 tatccgcggt accgagcacc atgtcgaatc tgagcaaagg cacgggcagc cggaaggaca 841 ccaagatgcg gatccgggcc tttccgatga ccatggatga aaaatatgta aacagcattt 901 gggaccttct gaaaaatgca attcaagaaa tccagcgtaa gaataacagt ggtcttagtt 961 ttgaggagct ctatagaaat gcatatacaa tggttttgca taaacatgga gaaaagctct 1021 acactggact aagagaagtt gttaccgaac atctcataaa taaggtgcga gaagatgtac 1081 taaattcatt gaataacaac tttcttcaaa cgctaaatca agcttggaat gatcatcaaa 1141 cagctatggt gatgattaga gacatactaa tgtacatgga ccgtgtgtat gtacaacaaa 1201 ataatgtgga gaacgtctac aatttgggat taattatttt tcgagatcaa gttgtacgtt 1261 atgggtgtat tagggatcat ctacggcaaa ctctattgga tatgattgca agagagcgga 1321 aaggagaagt cgtagacaga ggcgcaataa gaaatgcttg ccagatgtta atgattttag 1381 gtctcgaagg aagatcagtc tgtgaagaag attttgaggc tccttttttg gaaatgtctg 1441 cagaattttt tcagatggaa agccagaaat ttttagcaga aaatagtgct tcagtatata 1501 taaagaaagt agaagctaga attaatgaag aaatagaacg agtgatgcac tgccttgaca 1561 aatcaacgga agaaccaatt gtaaaggtgg ttgaaaggga actcatttcc aagcacatga 1621 agactatagt agaaatggag aattctgggc tagtacatat gttgaaaaat ggaaagacag 1681 aagaccttgg ttgcatgtac aagttattta gtcgtgtgcc aaatggtttg aaaacaatgt 1741 gtgagtgtat gagttcctat ttgagggagc aaggtaaagc tcttgtttct gaagaaggag 1801 aaggaaagaa tcctgttgac tatatccagg gcttattgga tctgaagagt aggttcgatc 1861 gcttcctcct ggaatcattc aacaatgacc gtctctttaa acaaactatt gcgggtgact 1921 ttgagtattt tctcaacctc aactccaggt ctcctgaata cctctcatta tttattgatg 1981 ataagctgaa aaagggagtc aaagggctaa cagaacaaga agtagaaaca atattggata 2041 aagcaatggt cctttttagg tttatgcaag aaaaagatgt atttgaacgt tattataaac 2101 aacacttggc aaggagactt ctcacaaata aaagtgtttc tgatgactct gaaaaaaaca 2161 tgatatctaa gttaaagact gaatgtggat gtcagttcac gtcaaaactg gaaggaatgt 2221 ttagggatat gagcatctca aacacaacga tggatgaatt caggcaacat ctacaggcaa 2281 ctggtgtatc tttaggtggt gttgatctta cagtccgggt gctcacgaca ggatattggc 2341 ccactcagtc agccacacca aagtgcaaca tcccaccagc accaagacat gcttttgaga 2401 tattcagaag gttctactta gccaaacaca gtggtcgaca gctcacactc cagcatcata 2461 tgggttctgc agatctcaat gccacatttt atggaccagt taaaaaggaa gatggatctg 2521 aagttggtgt tggaggtgca caagtaactg gctctaatac acggaagcac atattgcaag 2581 tttccacttt ccagatgacc atattaatgc tctttaataa tagagaaaaa tacacatttg 2641 aggaaattca gcaagagaca gatatccctg aaagagagct tgttagagcc ctacagtccc 2701 tcgcctgtgg taaaccaaca cagcgggttc ttacaaaaga acccaaatca aaggaaatag 2761 aaaatggtca tatatttaca gttaatgatc aattcacatc caaactacac agagtcaaga 2821 ttcaaacagt tgctgccaaa caaggtgaat ccgacccaga gaggaaagaa acaaggcaga 2881 aagtagacga cgacagaaaa catgagatag aagctgctat agtgcggata atgaaatcta 2941 gaaagaagat gcagcacaat gttctagtag cggaggtaac tcagcagttg aaggcgcgat 3001 tcttaccaag tccagttgtt attaagaaac gtattgaagg acttattgag agagaatatt 3061 tggcacgaac acctgaggat cgcaaagtat acacatatgt agcataaaat gcgttcagaa 3121 atttgattta ttcttggact gtactcttcg catggactgg gaagttcttt taaatcatta 3181 aatattaaga cgaccatctc ttctattaaa ttacagtaca tgttctagac cattgagatc 3241 aagcctttac tccctttgag agtttccaac atcagttgat tgagcttcag gctttacaac 3301 gtttatccct gtagagatca tctttacagt tcctcgggaa aatgtgaatg tgctgcgttt 3361 tgttttcttt actgtatgaa aacaggaaaa taaaagagaa atttagaaaa tacagctcat 3421 tacaataaaa ttgttggatt tcatttcaaa aaaaaaaaaa aaaaaactcg agcatgcatc 3481 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg 3541 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 3601 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 3661 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 3721 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa 3781 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg 3841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 3901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 3961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 4021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 4081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 4141 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 4201 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt 4261 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 4321 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 4381 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 4441 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 4501 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg 4561 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca 4621 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 4681 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 4741 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 4801 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 4861 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 4921 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 4981 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 5041 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 5101 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 5161 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 5221 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 5281 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 5341 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 5401 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa 5461 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 5521 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 5581 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 5641 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 5701 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 5761 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 5821 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 5881 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 5941 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 6001 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 6061 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 6121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 6181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 6241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 6301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 6361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 6421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 6481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 6541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 6601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 6661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 6721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 6781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 6841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 6901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 6961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 7021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 7081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 7141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 7201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 7261 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 7321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 7381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 7441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 7501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 7561 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 7621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 7681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 7741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 7801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 7861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 7921 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc 7981 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt 8041 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg 8101 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg 8161 ccagatatac gcgtt //