Gene:
Gene Symbol:
AF1265
Gene Name:
metal-dependent hydrolase
Original Clone ID:
3089g2BSt6b2
Keyword:
None
Species:
Archaeoglobus fulgidus
Type:
cDNA
Vector Name:
pSGX4
Format:
FUSION
Source:
New York SGX Research Center for Structural Genomics
Description:
None
Comments:
"N-terminal 6-His and Smt3, cleavable, TOPO"
Mutation / Discrepancy:
No / Yes
Publications:
None
Authors:
PSI
New York SGX Research Center for Structural Genomics
New York SGX Research Center for Structural Genomics
AsCD00311915
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AVAILABLE
No restriction
Special MTA: PSI/Smt3/SUMO
Special MTA: PSI/Smt3/SUMO
Insert sequence: 687nts
Open reading frame: 51 to 737
AGGATAACGATATTATTGAGGCTCACAGAGAACAGATTGGTGGATCCCTTAAAATAACAT
GGTTGGGACACGCGGCCTTTTTGCTTGAGGGAAGCATGAAGGTGCTGATTGACCCCTTCC
TCACGGGAAATCCCAACGCTCCCGTAAAGCCTGAAGAAGTCAAGGCGGATTACATCCTCG
TAACCCACGGCCACGGAGACCATCTTGGAGATGCGGTTGAGATTGCCAAGAGAAACAACG
CCCCCATAATCTGCATTCACGAGCTTTCGAGAATTCTTTCGAGGTATGATGTCGAAACTA
TGGGAATGAACATGGTTGGGACTGCGAGAACGGGAAGCATTGCAGTTACAATGGTTCCAG
CCTGGCATTCAGCAGATTTGGAAGACGAGCAGGGGAACATAATAAGCGCTGGCCTTCCAG
CGGGCTTTATCGTCTCAATGGATGGCGTGAGAGTTTACCACACTGGCGACACGGATGTCT
TTCTGGACATGCAGCTAATCGGAGAGCTCCACAGGCCAGATGTGATGCTTCTGCCCATCG
GAGACTACTACACCATGGGTATTGCCGGAGCCGTTAAGGCTTTGGAGCTTGTTAAGCCCA
AAGTAGCGATACCGATGCACTACAACACCTTCCCGCTGGTCGAGAAGGACCCAGAGGACT
TCAGAAAGGCTGTGAAGGCGAAGGGGCTGGATGTCGAAGTTGTGATTCTAAAGCCGGGAG
AGAGCTACGAGTTCAACAAAGGGTAACTCGAGCACCACCACCACCACCACGAGATCCGGT
GTAACAAG
Protein sequence predicted based on nucleotide sequence
KITWLGHAAFLLEGSMKVLIDPFLTGNPNAPVKPEEVKADYILVTHGHGDHLGDAVEIAK RNNAPIICIHELSRILSRYDVETMGMNMVGTARTGSIAVTMVPAWHSADLEDEQGNIISA GLPAGFIVSMDGVRVYHTGDTDVFLDMQLIGELHRPDVMLLPIGDYYTMGIAGAVKALEL VKPKVAIPMHYNTFPLVEKDPEDFRKAVKAKGLDVEVVILKPGESYEFN
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Mutations:
None
Discrepancy:
g266t,G89V,del689-691
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:
ProteinExpressed: Protein_Confirmed
SolubleProtein: Protein_Soluble
ProteinPurified: Tested_Not_Purified
Find experimental protocols in NYSGXRC-3089g
Recommended expression in:
Not Applicable
Vector Name:
pSGX4
Synonyms:
pSGX-4, pSGX4_BS
Sequencing Primer:
Forward:
T7
Reverse:
T7 terminator
Description:
Bacterial (E. coli) expression vector with T7 promotor, adds N-terminal 6xHis and smt-3 tags, cleavage site; kanamycin resistance; TOPO cloning.
Comments:
Modified from pET26b from Novagen. N-terminal 6xHis-tag followed by smt3 fusion protein. Smt3 fusion cloned into NheI and BamHI sites. TOPO adaptation at BamHI and XhoI sites.
Size (bp):
5611
Parent Vector:
None
Properties:
TOPO Cloning, bacterial expression, in vitro transcription, with tag/fusion/marker
Author Name:
New York SGX Research Center for Structural Genomics
Publications:
None
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
TOPO recognition site
TOPO
TOPO cloning site
5435
5444
bacterial origin
ColE
ColE1 bacterial origin of replication
1421
2103
gene fragment
ROP
ROP
2527
2703
promoter
T7
T7 promoter
4984
5003
protease cleavage site
ULP site
ULP cleavage site (recognizes the tertiary structure of SUMO)
5140
5140
repressor protein gene
LacI
lacI cds
3515
4351
ribosome binding site
RBS
ribosome binding site (RBS)
5062
5068
selectable marker
KanR
kanamycin resistance gene
563
1375
tag
Smt3
Smt3 tag
5140
5424
tag
His
6xHis tag
5083
5100
trxn regulatory element
lac operator
lac operator
5003
5025
trxn termination sequence
T7 term
T7 termination sequence
5539
5585
