质粒名称: | pCDEF3-vhs |
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出品公司: | 爱迪基因 |
目录编号: | 131696 |
存储实验室: | Britt Glaunsinger |
载体骨架基本信息 |
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载体名称: | pCDEF |
载体抗性: | 氨苄青霉素 |
完整质粒大小: | 6776 bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | vhs |
物种来源: | HSV-1 |
插入大小: | 1469 bp |
启动子 | CMV |
5' 测序引物 | CGCAAATGGGCGGTAGGCGTG |
3' 测序引物 | gcgatgcaatttcctcattt |
文献引用方法 |
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材料和方法部分: | pCDEF3-vhs was a gift from Britt Glaunsinger (Addgene plasmid # 131696 ; http://n2t.net/addgene:131696 ; RRID:Addgene_131696) |
买家导航
pCDEF3vhs载体基本信息
pCDEF3vhs载体图谱质粒图谱和多克隆位点信息
pCDEF3vhs载体简介
pCDEF3vhs载体序列
LOCUS pCDEF3-vhs 7130 bp ds-DNA circular SYN 02-OCT-2019 DEFINITION expresses vhs in mammalian cells. ACCESSION . VERSION . KEYWORDS pCDEF3-vhs SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7130) TITLE Xrn1 activity broadly represses RNA polymerase II occupancy at mammalian but not viral promoters during herpesvirus infection JOURNAL Unpublished REFERENCE 2 (bases 1 to 7130) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 2, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7130 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(43..62) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 234..613 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 614..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 768..788 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 862..881 /label=T7 /note="T7 promoter, forward primer" promoter 862..880 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2720..2737) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2726..2950 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2996..3424 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3083..3102) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3293..3314 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3433..3453) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3438..3767 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3618..3753 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3680..3699 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3834..4628 /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(3888..3907) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4498..4517 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4802..4923 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4839..4858) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4893..4912 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4972..4988) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4972..4988) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4985..5007) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4996..5012 /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(5020..5050) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5065..5086 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5203..5220) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5374..5962) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5454..5473) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6133..6993) /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 6756..6775 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6994..7098) /gene="bla" /label=AmpR promoter ORIGIN 1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc 61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc 121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct 181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg 241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat 301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat 601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga 661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg 781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac 841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg 901 tttaaactta agcttggtac cgagctcgga catgggtttg ttcgggatga tgaagtttgc 961 ccacacacac catctggtca agcgccaggg ccttggggcc ccggccgggt acttcacccc 1021 catcgccgtg gacctgtgga acgtcatgta cacgttggtg gtcaaatatc agcgccgata 1081 ccccagttac gaccgcgagg ccattacgct acactgcctc tgtcgcttat taaaggtgtt 1141 tacccaaaag tcccttttcc ccatcttcgt taccgatcgc ggggtcaatt gtatggagcc 1201 ggttgtgttt ggagccaagg ccatcctggc ccgcacgacg gcccaatgcc ggacggacga 1261 ggaggccagt gacgtggacg cctctccacc gccttccccc atcaccgact ccagacccag 1321 ctctgccttt tccaacatgc gccggcgcgg cacctctctg gcctcgggga cccgggggac 1381 ggccgggtcc ggagccgcgc tgccgtccgc cgcgccctcg aagccggccc tgcgtctggc 1441 gcatctgttc tgtattcgcg ttctccgggc cctggggtac gcctacatta actcgggtca 1501 gctggaggcg gacgatgcct gcgccaacct ctatcacacc aacacggtcg cgtacgtgta 1561 caccacagac actgacctcc tgttgatggg ctgtgatatt gtgttggata ttagcgcctg 1621 ctacattccc acgatcaact gtcgcgatat actaaagtac tttaagatga gctaccccca 1681 gttcctggcc ctctttgtcc gctgccacac cgacctccat cccaataaca cctacgcctc 1741 cgtggaggat gtgctgcgcg aatgtcactg gacccccccg agtcgctctc agacccggcg 1801 ggccatccgc cgggaacaca ccagctcgcg ctccacggaa accaggcccc ctttgccgcc 1861 ggccgccggc ggcaccgaga tgcgcgtctc gtggaccgaa attctaaccc aacagatcgc 1921 cggcggatac gaagacgacg aggacctccc cctggatccc cgggacgtta ccgggggcca 1981 ccccggcccc aggtcgtcct cctcggagat actcaccccg cccgagctcg tccaggtccc 2041 gaacgcgcag ctgctggaag agcaccgcag ttatgtggcc agccggcgac gccacgtcat 2101 ccacgacgcc ccagagtccc tggactggct ccccgatccc atgaccatca ccgagctggt 2161 ggaacaccgc tacattaagt acgtcatatc gcttatcggc cccaaggagc gggggccgtg 2221 gactcttctg aaacgcctgc ctatctacca ggacatccgc gacgaaaacc tggcgcgatc 2281 tattgtgacc cggcatatca cggcccctga tatcgccgac aggtttctgg agcagctgcg 2341 gacccaggcc cccccacccg cgttctacaa ggacgtcctg gccaaattct gggacgagta 2401 gcccaaacgt cagacgagcg cgcttgtccc cgaacaaacg acccaccaat aaaattatgg 2461 tatcctatgc cctcagaatc tggacggacc tggttactgc tttttgcgcc gccttttatc 2521 ctctcccacc cccgcgtccc tgacaagaat cacaatgaga cccaaagttt ggttcagagg 2581 tttattatgg gcaaacacgg gtagaagcgc gccgcgaccc tcacagatcg ttagcttggt 2641 accgagctcg aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc 2701 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 2761 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 2821 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 2881 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 2941 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg 3001 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3061 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3121 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 3181 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 3241 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 3301 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 3361 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3421 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3601 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3661 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3721 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3781 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3841 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3901 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3961 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 4021 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 4081 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 4141 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 4201 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 4261 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 4321 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 4381 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 4441 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 4501 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 4561 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4621 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 4681 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 4741 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 4801 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 4861 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4921 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 4981 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 5041 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 5101 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 5161 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 5221 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 5281 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 5341 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 5401 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 5461 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 5521 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 5581 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 5641 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 5701 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5761 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 5821 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 5881 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 5941 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 6001 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 6061 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 6121 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 6181 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 6241 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 6301 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 6361 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 6421 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 6481 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 6541 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 6601 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 6661 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 6721 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 6781 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 6841 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 6901 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 6961 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 7021 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 7081 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg //
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