Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
1038.1.24_GO.33429
Keyword:
None
Species:
Arabidopsis thaliana Columbia
Type:
cDNA
Vector Name:
pVP16
Format:
CLOSED
Source:
Center for Eukaryotic Structural Genomics
Description:
None
Comments:
His8-MBP fusion prtotein 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
AtCD00085119
Price:
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 975nts
Open reading frame: 28 to 1002
ACAAGTTTGTACAAAAAAGCAGGCTCCGAAAACCTGTACTTCCAGTCCGCGTCTCTCAAA
GTCCCAAGCAATGTTCCTCTTCCCGAAGATGACGCCGAGCAACTCCACAAGGCTTTTTCA
GGATGGGGTACCAACGAGAAGCTGATCATATCAATACTAGCTCACAGGAACGCAGCACAA
CGCAGCTTGATCCGCAGCGTTTATGCAGCTACCTACAATGAGGATCTTCTCAAAGCATTA
GACAAAGAGCTTTCTAGCGACTTTGAGAGAGCTGTGATGTTGTGGACTCTTGATCCACCA
GAGAGAGATGCTTATTTGGCTAAAGAATCCACCAAGATGTTCACCAAGAACAATTGGGTT
CTTGTTGAAATCGCTTGCACAAGGCCTGCTCTTGAGCTTATCAAGGTCAAGCAAGCTTAC
CAAGCTCGATACAAGAAATCAATCGAGGAAGATGTCGCGCAACACACATCTGGTGACCTT
CGTAAGCTCTTGCTTCCTCTTGTGAGCACTTTCAGGTATGAAGGAGATGATGTGAACATG
ATGCTTGCAAGATCTGAAGCTAAGATACTTCACGAGAAGGTCTCAGAGAAATCTTACAGT
GACGATGACTTCATCAGAATCTTGACAACAAGAAGCAAAGCACAGCTCGGTGCAACACTC
AACCACTACAACAACGAGTATGGAAACGCCATTAACAAGAACTTGAAGGAAGAGTCGGAC
GACAATGACTACATGAAACTACTAAGAGCTGTAATCACATGTTTGACATACCCTGAGAAG
CATTTTGAGAAGGTTCTTCGTCTATCAATCAACAAAATGGGAACAGACGAATGGGGACTA
ACCCGAGTCGTGACTACACGAACTGAAGTTGACATGGAACGCATCAAAGAGGAATATCAG
CGAAGAAACAGCATTCCTTTGGACCGTGCTATCGCCAAAGACACTTCTGGTGACTATGAG
GACATGCTTGTTGCTCTTCTCGGACATGGCGATGCTTGATAGGACCCAGCTTTCTTGTAC
AAAGTGGT
Protein sequence predicted based on nucleotide sequence
ENLYFQSASLKVPSNVPLPEDDAEQLHKAFSGWGTNEKLIISILAHRNAAQRSLIRSVYA ATYNEDLLKALDKELSSDFERAVMLWTLDPPERDAYLAKESTKMFTKNNWVLVEIACTRP ALELIKVKQAYQARYKKSIEEDVAQHTSGDLRKLLLPLVSTFRYEGDDVNMMLARSEAKI LHEKVSEKSYSDDDFIRILTTRSKAQLGATLNHYNNEYGNAINKNLKEESDDNDYMKLLR AVITCLTYPEKHFEKVLRLSINKMGTDEWGLTRVVTTRTEVDMERIKEEYQRRNSIPLDR AIAKDTSGDYEDMLVALLGHGDA**
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Reference Sequence Annotations:
Start on reference sequence
1
End on reference sequence
975
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:
ProteinExpressed: Protein_Confirmed
ProteinPurified: Protein_Purified
SolubleProtein: Protein_Soluble
Find experimental protocols in GO.33429
Recommended expression in:
Not Applicable
Vector Name:
pVP16
Synonyms:
''
Sequencing Primer:
Forward:
MBP Forward
Reverse:
pQE Reverse
Description:
Bacterial (E. coli) expression, adds N-terminal 8xHis 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):
7631
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
6471
5789
negative selection marker
ccdB
ccdB death cassette gene (lost in with-insert form)
2643
2852
promoter
T5
T5 promoter
5
54
recombination site
AttR1
AttR1 recombination site
1316
1440
recombination site
AttR2
AttR2 recombination site
3020
2896
repressor protein gene
LacI
LacI coding region
4132
5211
selectable marker
CmR
chloramphenicol resistance gene (CmR) (Lost in with-insert form)
1549
2205
selectable marker
AmpR
ampicillin resistance gene
7228
6569
tag
MBP
N-terminal MBP
148
1246
tag
His
N-terminal 8xHis tag
128
145
