Gene:
Gene Symbol:
Mrad2831_3263
Gene Name:
enoyl-CoA hydratase/isomerase
Original Clone ID:
28397
Keyword:
None
Species:
Methylobacterium radiotolerans JCM 2831
Type:
cDNA
Vector Name:
pNIC28-Bsa4
Format:
CLOSED
Source:
New York Structural Genomics Research Consortium
Description:
None
Comments:
None
Mutation / Discrepancy:
No / No
Publications:
None
Authors:
Albert Einstein College of Medicine
Steven Almo
PSI:Biology
New York Structural Genomics Research Consortium
Steven Almo
PSI:Biology
New York Structural Genomics Research Consortium
MrCD00598620
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 801nts
Open reading frame: 24 to 824
AGAACCTGTACTTCCAATCCATGAGTGCCGAGACTGGGACCCGCGACGCGGGCGAGGACC
TGTTGTTCTCCGTCGCCGAGGGGGTCGCGACCATTACGCTGAACCGGCCCGAGCGGAAGA
ACGCCTTCACCTTCCCGATGATCGAGGCCTGGACGGCGGCCCTGCAGCGCTGCCGCACCG
ACGCGGCGATCCGCGTCGTGGTGGTGACCGGCGCGGGCTCGGCCTTCTGCAGCGGCGGCG
ACATCGTCGAGATGGGGGAGCGCCTCGACCAGCCGCCGGCCCGGCGCAAGGAGGAGCTCC
ACGGGCGGATCCAGCGCATCCCGCTGACGCTGGAGGATCTCGACAAGCCGGTGATCGCCG
CCCTGAACGGCGTGGCCACCGGCGCGGGCCTCGACCTCGCCCTGATGTGCGACCTGCGCT
ACGCCGCGCAGAGCGCCCGCTTCGCCGAGACCTACGTGCGGGTCGGCTTGGTCCCGGGCG
CGGGCGGCGCCCATTTCCTGCCCCGGCTCGTCGGGACCGCGAAGGCGCTGGAGCTGTTCT
TCACCGGCGATTTCGTCGACGCCGACGAGGCCCTGCGGATCGGCCTCGTCAACGGCGTCC
ACGCCGACGCCGACCTGATGCCGGCCGTCGCGGCGCTGGCGCGGAAGATCGCCCAGGCGC
CGCCGCTGACCCTGAGCCTGATCAAGCGCGCGGTCTATCAGGGCATGCGCAACGACCTGC
GCACCAACCTCGACCTGATCTCCTCGCACTACGCCGTCGTCACCGCGACGCCCGAGCATC
GGGAGGCGGTGCAGGCCTTTATCGGCCAGCGCGACAGGGGCTAACAGTAAAGGTAA
Protein sequence predicted based on nucleotide sequence
SAETGTRDAGEDLLFSVAEGVATITLNRPERKNAFTFPMIEAWTAALQRCRTDAAIRVVV VTGAGSAFCSGGDIVEMGERLDQPPARRKEELHGRIQRIPLTLEDLDKPVIAALNGVATG AGLDLALMCDLRYAAQSARFAETYVRVGLVPGAGGAHFLPRLVGTAKALELFFTGDFVDA DEALRIGLVNGVHADADLMPAVAALARKIAQAPPLTLSLIKRAVYQGMRNDLRTNLDLIS SHYAVVTATPEHREAVQAFIGQRDRG*
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Mutations:
None
Discrepancy:
c40t,L14L
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:
kanamycin
Bacterial Selection Condition:
50ug/mL kanamycin
Growth Condition:
Growth with the single antibiotic in LB at 37 degrees is recommended.
Comments:
Conditions for Gateway-type vectors in recombined (with insert) form and other kanamycin resistant vectors.
Protein Expression Results:
SolubleProtein: Not_Applicable
ProteinPurified: Protein_Purified
ProteinExpressed: Tested_Not_Found
Find experimental protocols in NYSGRC-028397
Recommended expression in:
Not Applicable
Vector Name:
pNIC28-Bsa4
Synonyms:
pSEC-His
Description:
Bacterial expression vector with T7 promoter and N-terminal 6xHis tag with TEV protease cleavage site; kanamycin resistance in bacteria; ligation independent cloning
Comments:
pET expression vector with His6 tag in 22-aa N-terminal fusion peptide, with TEV protease cleavage site. Includes sites for LIC cloning, and a “stuffer” fragment that includes the SacB gene, allowing negative selection on 5% sucrose
Size (bp):
7284
Parent Vector:
None
Properties:
bacterial expression, ligation independent cloning (LIC), with tag/fusion/marker
Author Name:
Opher Gileadi
Publications:
PMID:
20541610
Title: High-throughput production of human proteins for crystallization: the SGC experience
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial origin
ColE
ColE1 pBR322 origin
4338
4338
bacterial origin
f1
f1 origin
2266
2721
ligation independent cloning
5'LIC
96
110
ligation independent cloning
3'LIC
2036
2050
primer
pLICfor
pLIC forward sequencing primer TGTGAGCGGATAACAATTCC
7262
7282
primer
pLICrev
pLIC reverse sequencing primer AGCAGCCAACTCAGCTTCC
2145
2164
promoter
T7
T7 promoter
7238
7254
protease cleavage site
TEV
TEV protease cleavage site
87
110
repressor binding site
lac operator
lac operator
7257
7281
repressor protein gene
LacI
Lac repressor gene
5772
6851
selectable marker
KanR
kanamycin resistance
2817
3629
selectable marker
SacB
SacB gene - allows for negative selection on 5% sucrose
569
1987
tag
His
N-terminal 6xHis tag
45
62
trxn termination sequence
T7 term
T7 terminator
2183
2229
