Gene:
Gene Symbol:
Gmet_0418
Gene Name:
radical SAM domain-containing iron-sulfur cluster-binding oxidoreductase
Original Clone ID:
GmR154-21.1
Keyword:
None
Species:
Geobacter metallireducens
Type:
cDNA
Vector Name:
pET21_NESG
Format:
FUSION
Source:
Northeast Structural Genomics Consortium
Description:
None
Comments:
None
Mutation / Discrepancy:
No / Yes
Publications:
None
Authors:
PSI
Northeast Structural Genomics Consortium
Northeast Structural Genomics Consortium
GmCD00546435
Price:
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AVAILABLE
No restriction
Special MTA: PSI/NESG
Special MTA: PSI/NESG
Insert sequence: 1671nts
Open reading frame: 51 to 1721
TCCCCCTCTAGAAAATAATTTTGTTTAACTTTAAGAAGGAGAATATACATATGTCGTGGA
AAATCATTGAAAAATACCGCCGTATCCTTGCCCAGGAAGAGGGGAATTCACCCAAGCGGC
GTGGCGGGAAACTGACCGTCTGCCTCGTCTATCCCAACCGCTATGGCGTGGCCATGAGCA
ACCTCGGCTTCCAGGCGGTCCATGCGCTGCTGAACGCCGAGTCCGACACCCTCTGTGAAC
GGGCTTTCCTCCCCGACCCGGACGATCTGCGGGAATACCGCCGCAGCGGCGCCCGGCTCC
TCTCCCTGGAGTCCCAGGAACCACTTTCCTCCTTCGATATTATTGCTTTTTCGGTTTCTT
TCGAGAGCGACTACCTGAACATCCCCGCCATCTTCGACCTGGCCGGGATCCCGGCCCTGG
CGGCTGAACGGACAGGGTCCCATCCCCTCGTCATGGGGGGCGGGGCGGTCCTCTTTCTGA
ATCCGGAGCCGGTGGCTGATTTCCTGGATCTCATTGTCGTGGGGGAGGGGGAGGCGATTC
TGCCGACGTTTCTGGACCACATTCGCGCCGACGGGATTGGGGACCGCGCAATGTTTCTTG
CAGGAGTCGCGGGGCTTCCCGGTATCTACGTGCCGTCTCTCTACGCGACGGAGTATGACG
GCGCCCGCCTGGTGTCGCGGGAGGCGAAGGACGGGGCGCCGGAGCAGGTGAAGCGTATCT
GGATGGTCGAGCCCGACGGGCGGGCCACGGTGTCCGAGATTTTGACGCCCGGCACCGAGT
TCTCCGACATGTATCTGGTTGAGGTCTCCCGGGGGTGCGGGCGGGGGTGCCGCTTCTGTG
CCGCCGGCTTCATCTACCATCCCTACCGGCAGCGCTCCCTCGACGCGGTGAAGGCCGCCG
TTGCCGAAGGACTCGGCCACCGGCGCAAGATCGGGCTCGTGGGGGCGGCGGTCTCGGATT
TTCGGGAGATCGGTGAACTCTGCCGGCACATCCTGGCGGAGGGGGGAACGGTGTCGGTGT
CGTCGCTGCGAATCGACCGCCTCGACGAGGGGATGATCGAGGTACTCAAGGCGAGCGGCC
ACAAGACCGTGGCCCTGGCCCCCGAGGGGGGGAGCCAGCGGATACGGGACCTCATCAACA
AGAACCTGACCGAGGAGCAGATTCTGGCCGCCTGCGATCTCCTCATCGGCCATGATATCC
TGAACCTGAAGCTCTACTTCATCATCGGCCTCCCCACCGAGACCATGGACGATCTGGAGG
AGATGGCCCTCCTCGTGGAGAAGATCCGCAAGCGGGTCATTGCCGCGGCGCGGAAAAACA
AGCGGCTCGGTGAGATCATCCTCTCCGTGAACCCCTTTGTCCCCAAGCCGTTCACACCGT
TCCAGTGGTGCGGCATGGAAGATACCCCCTCCCTAGAGAGAAAGCTCAAGTACCTCCAGC
AGACGGTGGGGAGGCTGTCAAACGTCCGGCTCATTGTCGAGAGCCCCAAGGATGCCTGCC
TCCAGGCCCTCCTCTCCCGAGGGGACCGGCGGCTCTCGGGGTTCCTCCTCCGCGCCGCGG
AACTGGGGAACTGGAAGCGGGCGGCCCGGGAATTTCCCGTCGACACCGACTTCCTGGTCC
ACCGGACCATCCCCCTCGACGAGATGCTGCCGTGGGATGTCATAGCCAGCGCCGACGCCG
AGCGGCTCAGGCGGGAGTATCTGAAGGCGACCCCTCCCCGG
Protein sequence predicted based on nucleotide sequence
MSWKIIEKYRRILAQEEGNSPKRRGGKLTVCLVYPNRYGVAMSNLGFQAVHALLNAESDT LCERAFLPDPDDLREYRRSGARLLSLESQEPLSSFDIIAFSVSFESDYLNIPAIFDLAGI PALAAERTGSHPLVMGGGAVLFLNPEPVADFLDLIVVGEGEAILPTFLDHIRADGIGDRA MFLAGVAGLPGIYVPSLYATEYDGARLVSREAKDGAPEQVKRIWMVEPDGRATVSEILTP GTEFSDMYLVEVSRGCGRGCRFCAAGFIYHPYRQRSLDAVKAAVAEGLGHRRKIGLVGAA VSDFREIGELCRHILAEGGTVSVSSLRIDRLDEGMIEVLKASGHKTVALAPEGGSQRIRD LINKNLTEEQILAACDLLIGHDILNLKLYFIIGLPTETMDDLEEMALLVEKIRKRVIAAA RKNKRLGEIILSVNPFVPKPFTPFQWCGMEDTPSLERKLKYLQQTVGRLSNVRLIVESPK DACLQALLSRGDRRLSGFLLRAAELGNWKRAAREFPVDTDFLVHRTIPLDEMLPWDVIAS ADAERLRREYLKATPPR
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Mutations:
None
Discrepancy:
t179c,I60T;t338c,V113A;
c419t,A140V;c677t,A226V;
a865g,S289G;g1014a,V338V;
g1365a,L455L;a1370g,K457R;
c1427t,A476V;g1449c,W483C
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:
ProteinPurified: Not_Tested
ProteinExpressed: Protein_Confirmed
SolubleProtein: Tested_Not_Soluble
Find experimental protocols in GmR154
Recommended expression in:
Not Applicable
Vector Name:
pET21_NESG
Synonyms:
pET21, pET21-modified, pET21Cc, pET21_NESG
Sequencing Primer:
Forward:
T7
Reverse:
T7 terminator
Description:
Bacterial (E. coli) expression vector with T7 lac promoter, adds C-terminal 6xHis tag; ampicillin resistance; restriction enzyme cloning.
Comments:
Modified Novagen pET vector-common MCS
Size (bp):
5415
Parent Vector:
None
Properties:
bacterial expression, in vitro transcription, multiple cloning site, with tag/fusion/marker
Author Name:
Gaetano T. Montleione
Thomas B. Acton
Northeast Structural Genomics Consortium
Thomas B. Acton
Northeast Structural Genomics Consortium
Publications:
PMID:
15808222
Title: Robotic cloning and Protein Production Platform of the Northeast Structural Genomics Consortium
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
MCS
MCS
MCS (includes SphI, XbaI, BamHI, EcoRI, XhoI)
157
253
bacterial origin
ori
pBR322-type bacterial origin of replication
3180
3862
gene fragment
ROP
ROP
2580
2756
primer
T7 forward
T7 forward sequencing primer
283
298
primer
T7 reverse
T7 reverse sequencing primer
66
84
promoter
T7
T7 promoter
280
299
repressor binding site
lacO
lacO regulatory sequence
258
280
repressor protein gene
LacI
LacI
752
1768
selectable marker
AmpR
ampicillin resistance gene
3960
4619
ssDNA origin
f1 ori
f1 origin
5081
5387
tag
His
C-terminal 6xHis tag
140
157
trxn termination sequence
T7 term
T7 termination sequence
68
72
