| 质粒名称: | pDONR223_PIK3CA_WT |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 81736 |
| 存储实验室: | Jesse Boehm, William Hahn, David Root |
|
载体骨架基本信息 |
|
| 载体名称: | pDONR223 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 大观霉素 |
| 空载体大小: | 2790 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | Gateway Entry载体 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PIK3CA,NM_006218.3 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | WT |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 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_WT was a gift from Jesse Boehm & William Hahn & David Root (Addgene plasmid # 81736 ; http://n2t.net/addgene:81736 ; RRID:Addgene_81736) |
| 参考文献部分: | Systematic Functional Interrogation of Rare Cancer Variants Identifies Oncogenic Alleles. Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn WC. Cancer Discov. 2016 Jul;6(7):714-26. doi: 10.1158/2159-8290.CD-16-0160. Epub 2016 May 4. 10.1158/2159-8290.CD-16-0160 PubMed 27147599 |
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_WT 5997 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_WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5997)
AUTHORS Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG,
Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
WC
TITLE Systematic Functional Interrogation of Rare Cancer Variants
Identifies Oncogenic Alleles.
JOURNAL Cancer Discov. 2016 Jul;6(7):714-26. doi:
10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
PUBMED 27147599
REFERENCE 2 (bases 1 to 5997)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5997
/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(3876..3975)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(3992..4011)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3993..4011)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4016..4032)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4016..4032)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 4463..5254
/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 5347..5935
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5836..5855
/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 atgatgcaca tcatggtggc tggacaacaa aaatggattg
3841 gatcttccac acaattaaac agcatgcatt gaacttgcca actttcttgt acaaagttgg
3901 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
3961 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
4021 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
4081 taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct cgacttcgct
4141 gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
4201 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
4261 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
4321 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
4381 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
4441 ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact
4501 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
4561 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
4621 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
4681 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
4741 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
4801 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
4861 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
4921 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
4981 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
5041 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
5101 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
5161 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
5221 gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc
5281 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
5461 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
5701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
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