Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
30460
Keyword:
None
Species:
Mesorhizobium loti
Type:
cDNA
Vector Name:
pSpeedET
Format:
CLOSED
Source:
Joint Center for Structural Genomics
Description:
None
Comments:
None
Mutation / Discrepancy:
No / Yes
Publications:
None
Authors:
PSI
Joint Center for Structural Genomics
Joint Center for Structural Genomics
MlCD00104787
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 900nts
Open reading frame: 51 to 950
CTGATAAAATTCATCATCATCATCATCACGAAAACCTGTACTTCCAGGGCATGTTTTTTG
AGACCACCTTCCATGACGCAACGGAGCAGGAAGCGTCCTTGCCAGGTTGGCGGCAGGAAT
ACCGACAGGTTTCGACCGGCGGCTATGACGGTCGAACCATGGCTATCCTGCTCCCAGGCG
TGGAGATTTTGCGCGAGACCATCAGTGTCGGCACGGAACAAATGTTCTCGGCGCCGGAAG
GCAACATGATTTTCTATTACTACCCTAAACGCGATCGAAGTGAATTGGTCCGCGGCGAAA
AAGCGACCGGCGTTTCAGGTTTCGCCTTAAACTGGACCAATCGCATCGGGTTCATGGACG
CCGATAGCGACCTGCTGATGCTCGTCCTAAGGCGCGATGCTGTTGCGGACGAGGACCGCA
GCAGAATAGCGGAGCTTGTCGAACCCCCTCTAGAGAATGCGGCTTTTCTTGCAGACTGGC
TGCTGTCGTTGCTTGGAGGCGCTCGCCAACAATCGCATCCGTGCGCCTGGGGAAAATTCG
ACGGCGTGCTCGCGGACGTCATTACAGATCGTTTCGAGCTGCTTTTCCAGAAAAGCCTCG
TTCGCCAAACGCCGCACATAGCGCAGACAGAACAAATCTATAGAAAGATCCGCGATCGAA
TTTTCGATGCGCCTGATATGCCCCATTCGGTGAAGTCGCTGTCGCGAGATCTTGGTGTAT
CGGCCTTCAACCTCAGGGCATCCTGCCAAAGTTTCACGCAGATCCCTCTCGACAGAATCC
TGATGATGCTTCGCTTGAACGGCGCCCGCCGCGACTTGCTGCACGCTCGTTGTTCGCCGC
GGAAGGTATCCGACATAGCAATGGATTGGGGCTTTTTTCACTGGAGCCGCTTTGCGCTGC
GCTACAAGTCGCTTTTCGGCGAAACACCCTCTCAGACCCTGAAGGGTTAACGCGACTTAA
TTAA
Protein sequence predicted based on nucleotide sequence
MFFETTFHDATEQEASLPGWRQEYRQVSTGGYDGRTMAILLPGVEILRETISVGTEQMFS APEGNMIFYYYPKRDRSELVRGEKATGVSGFALNWTNRIGFMDADSDLLMLVLRRDAVAD EDRSRIAELVEPPLENAAFLADWLLSLLGGARQQSHPCAWGKFDGVLADVITDRFELLFQ KSLVRQTPHIAQTEQIYRKIRDRIFDAPDMPHSVKSLSRDLGVSAFNLRASCQSFTQIPL DRILMMLRLNGARRDLLHARCSPRKVSDIAMDWGFFHWSRFALRYKSLFGETPSQTLKG*
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Reference Sequence Annotations:
Start on reference sequence
1
End on reference sequence
900
Mutations:
None
Discrepancy:
g194a,S65N;
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: Not_Applicable
ProteinPurified: Protein_Purified
SolubleProtein: Tested_Not_Soluble
Find experimental protocols in 373541
Recommended expression in:
Not Applicable
Vector Name:
pSpeedET
Synonyms:
pSpeed-ET
Sequencing Primer:
Forward:
pBAD forward
Reverse:
alt_bad_rev
Description:
Bacterial expression vector, arabinose or T7 induced expression, adds N-terminal MGSDKIHHHHHHENLYFQG tag; kanamycin resistance; PIPE cloning.
Comments:
If this vector contains an insert, the ccdB death cassette will not be present.
Size (bp):
5464
Parent Vector:
None
Properties:
PIPE cloning, bacterial expression, in vitro transcription, inducible expression, with tag/fusion/marker
Author Name:
Joint Center for Structural Genomics
Publications:
PMID:
18004753
Title: Combining the polymerase incomplete primer extension method for cloning and mutagenesis with microscreening to accelerate structural genomics efforts.
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial origin
ori
ColE1-type origin of replication
3306
3988
inducible trxn activator protein gene
araC
araC coding sequence
4563
5438
negative selection marker
ccdB
ccdB death cassette gene (lost in with-insert form)
1596
1805
promoter
T7
T7 promoter
290
310
promoter
araC
araC promoter
256
285
protease cleavage site
TEV
TEV protease cleavage site
382
402
selectable marker
CmR
chloramphenicol resistance gene (CmR) (lost in with-insert form)
502
1158
selectable marker
KanR
kanamycin resistance gene
2396
3208
tag
His
N-terminal 6xHis tag
364
381
trxn regulatory element
arabinose O1 operator
arabinose O1 operator
161
182
trxn regulatory element
arabinose O2 operator
arabinose O2 operator
5
23
trxn termination sequence
rrnB
rrnB T2 transcription terminator
2258
2284
