Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
ATCG3D_187.2
Keyword:
N-terminal and C-terminal deletion
Species:
Bacillus subtilis
Type:
cDNA
Vector Name:
pET28a
Format:
CLOSED
Source:
Accelerated Technologies Center for Gene to 3D Structure
Description:
None
Comments:
None
Mutation / Discrepancy:
No / No
Publications:
PMID: 19383143
Title: Combined protein construct and synthetic gene engineering for heterologous protein expression and crystallization using Gene Composer.
Title: Combined protein construct and synthetic gene engineering for heterologous protein expression and crystallization using Gene Composer.
Authors:
PSI
Accelerated Technologies Center for Gene to 3D Structure
Accelerated Technologies Center for Gene to 3D Structure
BsCD00430125
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 978nts
Open reading frame: 1 to 978
ATGGCTTCTATCAAAGTGATCGGTGTTGGCGGTGGCGGTAACAACGCGGTGAACCGTATG
ATTGAGAACGAAGTGCAAGGCGTTGAATACATCGCAGTGAACACCGATGCACAAGCTCTG
AACCTGTCCAAGGCTGAAGTTAAAATGCAAATCGGTGCTAAACTGACCCGTGGCCTGGGC
GCGGGCGCTAACCCGGAAGTTGGTAAGAAGGCGGCGGAGGAAAGCAAAGAACAGATCGAA
GAAGCCCTGAAAGGTGCTGACATGGTTTTCGTTACCGCGGGCATGGGTGGCGGTACTGGC
ACCGGCGCTGCACCTGTTATCGCTCAAATCGCTAAAGATCTCGGCGCTCTGACTGTCGGT
GTAGTTACTCGTCCTTTTACCTTCGAGGGTCGTAAACGTCAGCTGCAGGCTGCCGGTGGC
ATCTCCGCTATGAAAGAAGCGGTTGACACTCTGATCGTTATCCCGAACGACCGTATTCTG
GAGATCGTAGACAAGAATACCCCGATGCTGGAAGCATTCCGTGAAGCAGACAACGTTCTC
CGTCAGGGTGTTCAGGGCATCTCCGACCTTATCGCCACCCCGGGTCTGATCAACCTGGAC
TTCGCAGACGTAAAAACCATCATGTCTAACAAAGGCTCTGCTCTGATGGGCATTGGTATC
GCAACTGGTGAAAACCGCGCTGCAGAAGCAGCTAAGAAGGCGATTAGCTCCCCTCTGCTC
GAAGCGGCTATCGACGGCGCACAGGGTGTTCTGATGAACATTACCGGTGGTACCAACCTG
TCCCTTTACGAAGTTCAGGAAGCCGCTGACATCGTAGCGTCCGCTAGCGACCAAGATGTG
AACATGATCTTTGGCTCCGTTATCAACGAAAACCTTAAGGACGAAATTGTCGTAACCGTA
ATCGCTACCGGCTTCCTGGAAAACCTGTATTTTCAGGGCCATCATCATCATCATCATGAA
TATATGCCGATGGAATAA
Protein sequence predicted based on nucleotide sequence
MASIKVIGVGGGGNNAVNRMIENEVQGVEYIAVNTDAQALNLSKAEVKMQIGAKLTRGLG AGANPEVGKKAAEESKEQIEEALKGADMVFVTAGMGGGTGTGAAPVIAQIAKDLGALTVG VVTRPFTFEGRKRQLQAAGGISAMKEAVDTLIVIPNDRILEIVDKNTPMLEAFREADNVL RQGVQGISDLIATPGLINLDFADVKTIMSNKGSALMGIGIATGENRAAEAAKKAISSPLL EAAIDGAQGVLMNITGGTNLSLYEVQEAADIVASASDQDVNMIFGSVINENLKDEIVVTV IATGFLENLYFQGHHHHHHEYMPME*
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:
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
ProteinPurified: Protein_Purified
SolubleProtein: Protein_Soluble
Find experimental protocols in ATCG3D_187
Recommended expression in:
Not Applicable
Vector Name:
pET28a
Synonyms:
pET-28a, pET28
Sequencing Primer:
Forward:
T7
Reverse:
T7 terminator
Description:
Bacterial expression vector with T7lac promoter, adds N-terminal His tag, thrombin cleavage site, internal T7 epitope tag, C-terminal His tag; kanamycin resistance; restriction enzyme cloning.
Comments:
None
Size (bp):
5369
Parent Vector:
None
Properties:
bacterial expression, in vitro transcription, multiple cloning site, with tag/fusion/marker
Author Name:
Midwest Center for Structural Genomics
Publications:
None
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
MCS
MCS
MCS (XhoI, NotI, EagI, HindIII, SalI, SacI, EcoRI, BamHI, NheI, NdeI, NcoI)
158
203
bacterial origin
f1
f1 origin
4903
5358
bacterial origin
ori
bacterial origin of replication
3270
3889
promoter
T7
T7 promoter
368
386
repressor protein gene
LacI
lacI coding sequence (lacI cds)
896
1852
selectable marker
KanR
kanamycin resistance
3995
4807
tag
His
6xHis tag
140
157
trxn regulatory element
lac operator
lac operator
347
363
trxn termination sequence
T7 term
T7 terminator
26
72
