质粒名称: | pT/Caggs-NRASV12 |
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出品公司: | Addgene |
目录编号: | 20205 |
存储实验室: | John Ohlfest |
载体骨架基本信息 |
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载体名称: | colE1/ampR |
载体出品公司: | Ohlfest Lab |
空载体大小: | 5607 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 转座子 |
筛选标记: | -- |
载体抗性: | 氨苄霉素100 μg/mL |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 低拷贝 |
插入基因信息 |
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插入基因名称: | NRas |
物种来源: | H. sapiens (human) |
插入大小: | 570 bp |
基因突变: | G12V |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5’克隆位点: | -- |
3’克隆位点: | -- |
5’测序引物 | -- |
3’测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pT/Caggs-NRASV12 was a gift from John Ohlfest (Addgene plasmid # 20205 ; http://n2t.net/addgene:20205 ; RRID:Addgene_20205) |
参考文献部分: | De novo induction of genetically engineered brain tumors in mice using plasmid DNA. Wiesner SM, Decker SA, Larson JD, Ericson K, Forster C, Gallardo JL, Long C, Demorest ZL, Zamora EA, Low WC, SantaCruz K, Largaespada DA, Ohlfest JR. Cancer Res. 2009 Jan 15. 69(2):431-9. 10.1158/0008-5472.CAN-08-1800 PubMed 19147555 |
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pT/CaggsNRASV12载体基本信息
pT/CaggsNRASV12载体图谱质粒图谱和多克隆位点信息
pT/CaggsNRASV12载体简介
pT/CaggsNRASV12载体序列
LOCUS pT/Caggs-NRASV12 6300 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pT/Caggs-NRASV12 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6300) AUTHORS Wiesner SM, Decker SA, Larson JD, Ericson K, Forster C, Gallardo JL, Long C, Demorest ZL, Zamora EA, Low WC, SantaCruz K, Largaespada DA, Ohlfest JR TITLE De novo induction of genetically engineered brain tumors in mice using plasmid DNA. JOURNAL Cancer Res. 2009 Jan 15. 69(2):431-9. PUBMED 19147555 REFERENCE 2 (bases 1 to 6300) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6300 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..658 /label=chicken beta-actin promoter intron 659..1675 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1599..1621 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1683..1702 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" promoter complement(1748..1766) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1749..1766) /label=SP6 /note="SP6 promoter, forward primer" CDS 1800..1817 /codon_start=1 /product="epitope tag from middle T antigen of mouse polyoma virus" /label=Glu-Glu tag /translation="EYMPME" primer_bind complement(2395..2412) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2401..2625 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3079..3099) /label=T3 /note="T3 promoter, forward primer" promoter complement(3079..3097) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3118..3134) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3118..3134) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3131..3153) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3142..3158 /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(3166..3196) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3211..3232 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3349..3366) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3520..4108) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3600..3619) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4279..5139) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4902..4921 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5140..5244) /gene="bla" /label=AmpR promoter rep_origin 5271..5726 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5358..5377) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5568..5589 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5852..5874 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5866..5883 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 5867..5883 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 5893..5912 /label=T7 /note="T7 promoter, forward primer" promoter 5893..5911 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6285..1) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6286..2) /label=pBluescriptKS /note="For pBluescript vector" 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 gtcgaggtga gccccacgtt ctgcttcact ctccccatct 421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga 481 tgggggcggg gggggggggn ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg 541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc 601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg 661 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag 841 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt 901 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg 1021 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg 1081 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc 1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg 1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg 1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg 1321 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca 1381 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc 1441 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg 1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg 1561 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac 1621 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct 1681 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcc tcgagggggg 1741 gccctattct atagtgtcac ctaaatgcta gagctcgctg atccaccatg gaggaagagg 1801 aatatatgcc tatggaaatg actgagtaca aactggtggt ggttggagca gttggtgttg 1861 ggaaaagcgc actgacaatc cagctaatcc agaaccactt tgtagatgaa tatgatccca 1921 ccatagagga ttcttacaga aaacaagtgg ttatagatgg tgaaacctgt ttgttggaca 1981 tactggatac agctggacaa gaagagtaca gtgccatgag agaccaatac atgaggacag 2041 gcgaaggctt cctctgtgta tttgccatca ataatagcaa gtcatttgcg gatattaacc 2101 tctacaggga acagattaag cgagtaaaag actcggatga tgtacctatg gtgctagtgg 2161 gaaacaagtg tgatttgcca acaaggacag ttgatacaaa acaagcccac gaactggcca 2221 agagttacgg gattccattc attgaaacct cagccaagac cagacagggt gttgaagatg 2281 ctttttacac actggtaaga gaaatacgcc agtaccgaat gaaaaaactc aacagcagtg 2341 atgatgggac tcagggttgt atgggattgc catgtgtggt gatgtaagga tcagcctcga 2401 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 2461 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 2521 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 2581 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa 2641 gaaccagcca tcacaaagct ctgacctcaa tcctatagaa aggaggaatg agccaaaatt 2701 cacccaactt attgtgggaa gcttgtggaa ggctactcga aatgtttgac ccaagttaaa 2761 caatttaaag gcaatgctac caaatactaa ttgagtgtat gtaaacttct gacccactgg 2821 gaatgtgatg aaagaaataa aagctgaaat gaatcattct ctctactatt attctgatat 2881 ttcacattct taaaataaag tggtgatcct aactgaccta agacagggaa tttttactag 2941 gattaaatgt caggaattgt gaaaaagtga gtttaaatgt atttggctaa ggtgtatgta 3001 aacttccgac ttcaactgta tagggatcct ctagctagag tcgacctcga gggggggccc 3061 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc 3121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 3181 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 3241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 3301 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 3361 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 3421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 3481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 3541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 3601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 3661 gcttaccgga tacctgtccg cctttctccc ttcgggaacc gtggcgcttt ctcatagctc 3721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 3781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 3841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 3901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 3961 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 4021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 4081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 4141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 4201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 4261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 4321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 4381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 4441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 4501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 4561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 4621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 4681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaactt 4741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 4801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 4861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 4921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 4981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 5041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 5101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 5161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 5221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg 5281 tagcggcgca ttaagcgcgg cgggagtggt ggttacgcgc agcgtgaccg ctacacttgc 5341 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 5401 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 5461 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 5521 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 5581 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 5641 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 5701 taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa 5761 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca 5821 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc 5881 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg agctcggatc cctatacagt 5941 tgaagtcgga agtttacata cacttaagtt ggagtcatta aaactcgttt ttcaactact 6001 ccacaaattt cttgttaaca aacaatagtt ttggcaagtc agttaggaca tctactttgt 6061 gcatgacaca agtcattttt ccaacaattg tttacagaca gattatttca cttataattc 6121 actgtatcac aattccagtg ggtcagaagt ttacatacac taagttgact gtgcctttaa 6181 acagcttgga aaattccaga aaatgatgtc atggctttag aagctagatc tggccatcta 6241 gagcggccgc gcgcactagt gaattccatg gatatcaagc ttatcgatac cgtcgacctc //
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