Gene:
Gene Symbol:
AT4G14520
Gene Name:
DNA-directed RNA polymerase II-like protein
Original Clone ID:
5785.1.83_GO.16684
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
AtCD00334490
Price:
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 603nts
Open reading frame: 1 to 603
GAAAACCTGTACTTCCAGTCCGCTCGAGACGTGGCTATATGTGCCAAACACTTGAATGGC
CAATCGCCGCACCAACCGATTCTATGCAGACTACTCCAAGATCTAATACACGAGAAAGCG
TGCAGGGAACACGGATTCTATCTCGGCATCACCGCTCTAAAAAGCATCGGTAACAACAAG
AACAACAACATCGACAACGAAAACAATCATCAAGCAAAAATCCTCACATTCCCGGTGTCC
TTCACATGCCGCACATTCTTGCCTGCAAGAGGAGATATCTTGCAAGGGACTGTAAAAAAA
GTGTTGTGGAACGGAGCTTTTATCAGGTCAGGACCACTCAGATACGCATACCTCTCACTC
TTGAAAATGCCACATTATCATTATGTCCACTCACCATTGTCGGAGGACGAGAAACCGCAT
TTTCAGAAAGACGACCTCTCAAAGATTGCTGTTGGAGTTGTTGTGCGGTTTCAGGTGTTG
GCTGTGCGGTTTAAAGAGAGACCACACAAGAGAAGAAACGACTATTATGTTCTTGCAACT
TTGGAAGGCAATGGTTCATTTGGTCCCATCTCTCTTACAGGGTCAGATGAGCCATACATG
TAA
Protein sequence predicted based on nucleotide sequence
ENLYFQSARDVAICAKHLNGQSPHQPILCRLLQDLIHEKACREHGFYLGITALKSIGNNK NNNIDNENNHQAKILTFPVSFTCRTFLPARGDILQGTVKKVLWNGAFIRSGPLRYAYLSL LKMPHYHYVHSPLSEDEKPHFQKDDLSKIAVGVVVRFQVLAVRFKERPHKRRNDYYVLAT LEGNGSFGPISLTGSDEPYM*
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:
ProteinExpressed: Protein_Confirmed
SolubleProtein: Protein_Soluble
ProteinPurified: Tested_Not_Purified
Find experimental protocols in GO.16684
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
