Gene:
Gene Symbol:
AT4G22660
Gene Name:
putative F-box protein
Original Clone ID:
5784.1.86_GO.18961
Keyword:
full-length cds
Species:
Arabidopsis thaliana Columbia
Type:
cDNA
Vector Name:
pVP13
Format:
CLOSED
Source:
Center for Eukaryotic Structural Genomics
Description:
None
Comments:
His8-MBP fusion protein with TEV protease cleavage site is produced; after cleavage the protein obtained will be the translated CDS with the addition of SER at the N-term
Mutation / Discrepancy:
No / No
Publications:
None
Authors:
PSI
Center for Eukaryotic Structural Genomics
Center for Eukaryotic Structural Genomics
AtCD00334567
Price:
Login for Pricing
AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 1209nts
Open reading frame: 1 to 1209
GAAAACCTGTACTTCCAGTCCGAGAAGAACCAAAACCCTAATACCTGGTCGGATCTTCCT
CTAGATCTCTTGAACTTAGTTTTCAAACGCCTCAGCTTTGCTAATTTTCGACAAGCCAAA
TCGGTGTGTTCGTCTTGGTACTCTGCTTCGAAACAATCCGTGCCCAAAAATCAGATCCCT
TGGCTGATGCTCTTCCCCAAAGACAAAAACAACAACAAAAACAGTTCATGCACGATTTTT
TTCAATCCCGAGGACAAAGACCAACTTTACCAAACGCAAGATCTTGGTGTTGAATTTGCA
AAGAGTGTTTGTCTAGCAACCTATGGAAGTTGGCTCCTGATGCAAGATTCAAAGTATAAT
CTATACATTTTGAATCCATTCACCTACGAGAAGATCGGTCTTCCCGCTATTGAATCACAA
CAAGTTGGCATGGTAAAGGTTGACCAAACCATAGATGATGATTTTCTAACTTTCGACGAT
CACAACCACGTTAAGTTATTCAAAGGTAATAATACCGTACGTACCCCTGTGTTTTGGATT
GACGAGAAAACCAAAGAGTACATAGCTCTGTGGGGGCTTGGATATTGGTGTGTGGTTTAT
GCCAAGAACGGAGATAAGTTGTGGAATCAAATTCCAGAAATTATTTTAGATTCTCTTGAC
ATGGTTTACAAGGATCACAAGCTTTACTCATTTAGCTACAGAAATCTTTTCACAATCCTA
GATTTTTCTGGAGAGATTCCTCGAAAAGCCTTTCAATGTTTTATGCATGTCTATAGATCT
GAGTGGTTAAGTCCTCGTAGTCGTCAATTAAGTAATTCATGGTGCGTTGCTGAGACAAAA
CTTGTAGTCACAGTAACCGGAGATGTCCTCTTGGTTGAAAGAATGTTAAGACATTGGTCT
AGAATCCAGTCATTCAACGTCTACAAGTTTTATTCGTCAGGGACATTTCTTGATAAATAT
GAACTTGCTGATTCTTTGGGCGACGAGGCAATGCTTTTGGATCTAGGCATCACCGTGCTC
GCCAATGAAGTTGAGGGACTCCATAGAAATACTATATATTTCAGTGATAGCCATGACACG
ACCACAAAAGATCTCTTTCTGTTCAATTACGAGACACGGGAGATGGAGCCACTGCACAAA
TTTGATTTCGATTGTTCGTTACTTGAACTCTCTGCTCGATGGTTCTTACCAAGCTTTTCG
CATACATGA
Protein sequence predicted based on nucleotide sequence
ENLYFQSEKNQNPNTWSDLPLDLLNLVFKRLSFANFRQAKSVCSSWYSASKQSVPKNQIP WLMLFPKDKNNNKNSSCTIFFNPEDKDQLYQTQDLGVEFAKSVCLATYGSWLLMQDSKYN LYILNPFTYEKIGLPAIESQQVGMVKVDQTIDDDFLTFDDHNHVKLFKGNNTVRTPVFWI DEKTKEYIALWGLGYWCVVYAKNGDKLWNQIPEIILDSLDMVYKDHKLYSFSYRNLFTIL DFSGEIPRKAFQCFMHVYRSEWLSPRSRQLSNSWCVAETKLVVTVTGDVLLVERMLRHWS RIQSFNVYKFYSSGTFLDKYELADSLGDEAMLLDLGITVLANEVEGLHRNTIYFSDSHDT TTKDLFLFNYETREMEPLHKFDFDCSLLELSARWFLPSFSHT*
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Mutations:
None
Discrepancy:
None
5' Linker Sequence:
None
3' Linker Sequence:
None
Recommended Growth Condition:
Distributed in bacterial strain:
DH5-alpha T1 phage resistant
Antibiotic Selection:
Host Type:
bacterial
Marker:
ampicillin
Bacterial Selection Condition:
100 ug/mL ampicillin
Growth Condition:
Growth with the single antibiotic in LB at 37 degrees is recommended.
Comments:
Commonly used conditions for ampicillin resistant plasmid clones.
Protein Expression Results:
ProteinPurified: Not_Tested
SolubleProtein: Not_Tested
ProteinExpressed: Tested_Not_Found
Find experimental protocols in GO.18961
Recommended expression in:
Not Applicable
Vector Name:
pVP13
Synonyms:
''
Sequencing Primer:
Forward:
MBP Forward
Reverse:
pQE Reverse
Description:
Bacterial (E. coli) expression, adds N-terminal S-tag, 6xHis and MBP, adds TEV site on 5p end of insert; ampicillin resistance in bacteria; recombinational cloning; Gateway cloning into this vector will remove the Gateway ccdB death casette.
Comments:
TEV protease cleavage site is "partial" and not functional. The TEV cleavage site must be added to the insert by PCR, which allows amino acids encoded by the attB1 site to be cut off after protein expression. Gateway cloning into this vector will remove the Gateway cassette
Size (bp):
7675
Parent Vector:
None
Properties:
Gateway, acceptor (destination), bacterial expression, recombinational cloning, with tag/fusion/marker
Author Name:
Center for Eukaryotic Structural Genomics
Publications:
PMID:
15750721
Title: Results from high-throughput DNA cloning of Arabidopsis thaliana target genes using site-specific recombination
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial origin
ori
ColE1-type bacterial origin of replication
6516
5834
negative selection marker
ccdB
ccdB death cassette gene (lost in with-insert form)
2688
2897
promoter
T5
T5 promoter
7
87
recombination site
AttR1
AttR1 recombination site
1361
1485
recombination site
AttR2
AttR2 recombination site
3065
2941
repressor binding site
LacI
LacI coding sequence
4177
5256
selectable marker
CmR
chloramphenicol resistance gene (CmR) (lost in with-insert form)
1594
2250
selectable marker
AmpR
ampicillin resistance gene
7273
6614
tag
S-tag
N-terminal S-tag
116
152
tag
MBP
N-terminal MBP tag
193
1291
tag
His
N-terminal 6xHis tag
173
190
