| 质粒名称: | pDONR223_PIK3CA_p.H1047R |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 82824 |
| 存储实验室: | Jesse Boehm, Matthew Meyerson, David Root |
|
载体骨架基本信息 |
|
| 载体名称: | pDONR223 |
| 载体出品公司: | Invitrogen |
| 完整质粒大小: | 6000 bp |
| 载体类型: | Gateway Entry vector |
| 载体抗性: | Spectinomycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PIK3CA / NM_006218 |
| 物种来源: | H. sapiens (human) |
| 基因突变: | H1047R |
| 启动子: | None |
| 克隆方法: | Gateway Cloning |
| 5’测序引物 | M13pUC-fwd |
| 3’测序引物 | M13-Rev |
|
文献引用方法 |
|
| 材料和方法部分: | pDONR223_PIK3CA_p.H1047R was a gift from Jesse Boehm & Matthew Meyerson & David Root (Addgene plasmid # 82824 ; http://n2t.net/addgene:82824 ; RRID:Addgene_82824) |
| 参考文献部分: | High-throughput Phenotyping of Lung Cancer Somatic Mutations. Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS. Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28. 10.1016/j.ccell.2016.06.022 PubMed 27478040 |
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 pDONR223_PIK3CA_p.H1047R 6000 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gateway Donor vector containing PIK3CA, part of the Target
Accelerator Plasmid Collection..
ACCESSION .
VERSION .
KEYWORDS pDONR223_PIK3CA_p.H1047R
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6000)
AUTHORS Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F,
Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE,
Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B,
Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
TITLE High-throughput Phenotyping of Lung Cancer Somatic Mutations.
JOURNAL Cancer Cell. 2016 Aug 8;30(2):214-28. doi:
10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
PUBMED 27478040
REFERENCE 2 (bases 1 to 6000)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6000
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 92..109
/label=L4440
/note="L4440 vector, forward primer"
terminator 268..295
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 522..544
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 536..553
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(3879..3978)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(3995..4014)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3996..4014)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4019..4035)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4019..4035)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 4466..5257
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 5350..5938
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5839..5858
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
661 agttggcacc atgcctccac gaccatcatc aggtgaactg tggggcatcc acttgatgcc
721 cccaagaatc ctagtagaat gtttactacc aaatggaatg atagtgactt tagaatgcct
781 ccgtgaggct acattaataa ccataaagca tgaactattt aaagaagcaa gaaaataccc
841 cctccatcaa cttcttcaag atgaatcttc ttacattttc gtaagtgtta ctcaagaagc
901 agaaagggaa gaattttttg atgaaacaag acgactttgt gaccttcggc tttttcaacc
961 ctttttaaaa gtaattgaac cagtaggcaa ccgtgaagaa aagatcctca atcgagaaat
1021 tggttttgct atcggcatgc cagtgtgtga atttgatatg gttaaagatc cagaagtaca
1081 ggacttccga agaaatattc tgaacgtttg taaagaagct gtggatctta gggacctcaa
1141 ttcacctcat agtagagcaa tgtatgtcta tcctccaaat gtagaatctt caccagaatt
1201 gccaaagcac atatataata aattagataa agggcaaata atagtggtga tctgggtaat
1261 agtttctcca aataatgaca agcagaagta tactctgaaa atcaaccatg actgtgtacc
1321 agaacaagta attgctgaag caatcaggaa aaaaactcga agtatgttgc tatcctctga
1381 acaactaaaa ctctgtgttt tagaatatca gggcaagtat attttaaaag tgtgtggatg
1441 tgatgaatac ttcctagaaa aatatcctct gagtcagtat aagtatataa gaagctgtat
1501 aatgcttggg aggatgccca atttgatgtt gatggctaaa gaaagccttt attctcaact
1561 gccaatggac tgttttacaa tgccatctta ttccagacgc atttccacag ctacaccata
1621 tatgaatgga gaaacatcta caaaatccct ttgggttata aatagtgcac tcagaataaa
1681 aattctttgt gcaacctacg tgaatgtaaa tattcgagac attgataaga tctatgttcg
1741 aacaggtatc taccatggag gagaaccctt atgtgacaat gtgaacactc aaagagtacc
1801 ttgttccaat cccaggtgga atgaatggct gaattatgat atatacattc ctgatcttcc
1861 tcgtgctgct cgactttgcc tttccatttg ctctgttaaa ggccgaaagg gtgctaaaga
1921 ggaacactgt ccattggcat ggggaaatat aaacttgttt gattacacag acactctagt
1981 atctggaaaa atggctttga atctttggcc agtacctcat ggattagaag atttgctgaa
2041 ccctattggt gttactggat caaatccaaa taaagaaact ccatgcttag agttggagtt
2101 tgactggttc agcagtgtgg taaagttccc agatatgtca gtgattgaag agcatgccaa
2161 ttggtctgta tcccgagaag caggatttag ctattcccac gcaggactga gtaacagact
2221 agctagagac aatgaattaa gggaaaatga caaagaacag ctcaaagcaa tttctacacg
2281 agatcctctc tctgaaatca ctgagcagga gaaagatttt ctatggagtc acagacacta
2341 ttgtgtaact atccccgaaa ttctacccaa attgcttctg tctgttaaat ggaattctag
2401 agatgaagta gcccagatgt attgcttggt aaaagattgg cctccaatca aacctgaaca
2461 ggctatggaa cttctggact gtaattaccc agatcctatg gttcgaggtt ttgctgttcg
2521 gtgcttggaa aaatatttaa cagatgacaa actttctcag tatttaattc agctagtaca
2581 ggtcctaaaa tatgaacaat atttggataa cttgcttgtg agatttttac tgaagaaagc
2641 attgactaat caaaggattg ggcacttttt cttttggcat ttaaaatctg agatgcacaa
2701 taaaacagtt agccagaggt ttggcctgct tttggagtcc tattgtcgtg catgtgggat
2761 gtatttgaag cacctgaata ggcaagtcga ggcaatggaa aagctcatta acttaactga
2821 cattctcaaa caggagaaga aggatgaaac acaaaaggta cagatgaagt ttttagttga
2881 gcaaatgagg cgaccagatt tcatggatgc tctacagggc tttctgtctc ctctaaaccc
2941 tgctcatcaa ctaggaaacc tcaggcttga agagtgtcga attatgtcct ctgcaaaaag
3001 gccactgtgg ttgaattggg agaacccaga catcatgtca gagttactgt ttcagaacaa
3061 tgagatcatc tttaaaaatg gggatgattt acggcaagat atgctaacac ttcaaattat
3121 tcgtattatg gaaaatatct ggcaaaatca aggtcttgat cttcgaatgt taccttatgg
3181 ttgtctgtca atcggtgact gtgtgggact tattgaggtg gtgcgaaatt ctcacactat
3241 tatgcaaatt cagtgcaaag gcggcttgaa aggtgcactg cagttcaaca gccacacact
3301 acatcagtgg ctcaaagaca agaacaaagg agaaatatat gatgcagcca ttgacctgtt
3361 tacacgttca tgtgctggat actgtgtagc taccttcatt ttgggaattg gagatcgtca
3421 caatagtaac atcatggtga aagacgatgg acaactgttt catatagatt ttggacactt
3481 tttggatcac aagaagaaaa aatttggtta taaacgagaa cgtgtgccat ttgttttgac
3541 acaggatttc ttaatagtga ttagtaaagg agcccaagaa tgcacaaaga caagagaatt
3601 tgagaggttt caggagatgt gttacaaggc ttatctagct attcgacagc atgccaatct
3661 cttcataaat cttttctcaa tgatgcttgg ctctggaatg ccagaactac aatcttttga
3721 tgacattgca tacattcgaa agaccctagc cttagataaa actgagcaag aggctttgga
3781 gtatttcatg aaacaaatga atgatgcacg tcatggtggc tggacaacaa aaatggattg
3841 gatcttccac acaattaaac agcatgcatt gaactgattg ccaactttct tgtacaaagt
3901 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
3961 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
4021 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
4081 agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc
4141 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
4201 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
4261 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
4321 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
4381 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
4441 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
4501 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
4561 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
4621 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
4681 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
4741 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
4801 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
4861 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
4921 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
4981 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
5041 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
5101 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
5161 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
5221 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa
5281 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
5341 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
5401 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
5461 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
5521 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
5581 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
5641 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
5701 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
5761 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
5821 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
5881 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
5941 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
