| 质粒名称: | pT/Caggs-NRASV12, pT3-CAGGS-NRasV12 |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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/Caggs-NRASV12载体 pT3-CAGGS-NRasV12质粒基本信息
pT/Caggs-NRASV12质粒图谱 pT3-CAGGS-NRasV12载体图谱和pT/Caggs-NRASV12载体序列 pT3-CAGGS-NRasV12质粒序列多克隆位点信息
pT/Caggs-NRASV12质粒 pT3-CAGGS-NRasV12载体简介
pT/Caggs-NRASV12质粒序列 pT3-CAGGS-NRasV12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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pT/Caggs-NRASV12质粒 pT3-CAGGS-NRasV12载体应用举例
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