| 载体基本信息 |
| 载体名称 |
pDONR223_PIK3CA_WT |
载体抗性 |
Spectinomycin |
| 载体长度 |
5997 bp |
载体类型 |
Gateway Entry vector |
| 拷贝数 |
High Copy |
启动子 |
None |
| 克隆方法 |
Gateway Cloning |
5引物 |
M13pUC-fwd |
| 3引物 |
M13-Rev |
pDONR223_PIK3CA_WT 载体载体图谱
pDONR223_PIK3CA_WT 载体载体序列
LOCUS Exported 5997 bp ds-DNA circular SYN 13-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
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date:
"2016-07"; volume: "6"; issue: "7"; pages: "714-26"
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"