Gene:
Gene Symbol:
Npun_R2539
Gene Name:
radical SAM domain-containing protein
Original Clone ID:
APC39020.2_SG261_pMCSG19B
Keyword:
partial cds
Species:
Nostoc punctiforme PCC 73102
Type:
cDNA
Vector Name:
pMCSG19B
Format:
CLOSED
Source:
Midwest Center for Structural Genomics
Description:
None
Comments:
None
Mutation / Discrepancy:
No / No
Publications:
None
Authors:
Jessica Bearden
PSI
Midwest Center for Structural Genomics
PSI
Midwest Center for Structural Genomics
OoCD00450795
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 1200nts
Open reading frame: 46 to 1245
AAACTCGGGGGATTGGGTACGAGACCTGTACTTCCATCCAATGCCCCTCTGGGTTTAGTA
ACAGTAGCCGCTATTCTGCCCCAAGAATGGGAGTTCAAGCTGGTAGATCGCAACATCCGC
GCAGCAACCGAAGAAGAATGGGCATGGGCAGATGTAGTAATTCTCTCTGCGATGATTGTC
CAGAAACAAGATTTACTTGAGCAAATCCAGGAAGCCAAAAAACGTGGCAAGTTAGTCGCA
GTCGGCGGTCCCTACCCCACCTCTGTACCTCACGAAGTTGAAAATGTTGGCGCAGATTTT
CTGATTTTGGATGAAGGGGAAATCACTTTGCCCATGTTTATTGAGGCAATTCAACGGGGT
GAAACTTCTGGAACTTTCCGCGCCACAGAAAAACCTGATGTCACAAGCACGCCAATACCC
CGCTTTGATTTATTAGAACTGAATGCTTATGACATGATGTCGGTGCAGTTTTCGCGCGGG
TGTCCCTTCCAGTGCGAATTTTGTGACATTATTGTTCTCTACGGGCGCAAACCACGCACC
AAAGCCCCAGCCCAACTTTTAGCAGAATTAGATTACCTCTATGAATTGGGTTGGCGGCGG
GGTGTGTTCATGGTGGATGACAACTTTATTGGCAACAAACGAAATGTAAAATTGTTGCTG
AAAGAGTTAAAAGTCTGGATGGCAGAACACAAGTATCCCTTCCGATTTGACACTGAAGCT
TCAATTGACTTGGCACAAGATGCAGAAATGTTGGAGTTGATGGTTGAGTGTGGTTTCTCG
GCGGTGTTTTTGGGAATCGAAACGCCGGATGAGGATAGTTTGCAAATGACCAAGAAGTTT
CAAAATACTCGCAGTTCTCTAACTGAGGCAGTGGAAACCATCATTAAAGCGGGATTGCGC
CCAATGGCTGGGTTTATTATCGGCTTTGATGGCGAAAAAGCAGGCGCAGGCGATCGCATT
GTCCGCTTTGCTGAACAAGCAGCTATTCCTTCTACTACCTTTGCAATGTTGCAAGCGTTA
CCTAACACCGCGCTTTGGCATCGCCTGAAAAAAGAAGGACGACTGCGGGAAAATCAAGAT
GGCAACATCAACCAGACAACATTGATGAACTTCTTGCCCACCCGTCCGCTAGAAGAACTT
GCACGAGAATATATTGAAGCATTTTGTACTTTATACGACCCAGTGCAGTACTTGGATCGT
ACCTATCGCTGTTTCTTGATGATGGGTTTGCCGAGTTGGAAATAACATTGGAAGTGGATA
ACGGTAA
Protein sequence predicted based on nucleotide sequence
PLGLVTVAAILPQEWEFKLVDRNIRAATEEEWAWADVVILSAMIVQKQDLLEQIQEAKKR GKLVAVGGPYPTSVPHEVENVGADFLILDEGEITLPMFIEAIQRGETSGTFRATEKPDVT STPIPRFDLLELNAYDMMSVQFSRGCPFQCEFCDIIVLYGRKPRTKAPAQLLAELDYLYE LGWRRGVFMVDDNFIGNKRNVKLLLKELKVWMAEHKYPFRFDTEASIDLAQDAEMLELMV ECGFSAVFLGIETPDEDSLQMTKKFQNTRSSLTEAVETIIKAGLRPMAGFIIGFDGEKAG AGDRIVRFAEQAAIPSTTFAMLQALPNTALWHRLKKEGRLRENQDGNINQTTLMNFLPTR PLEELAREYIEAFCTLYDPVQYLDRTYRCFLMMGLPSWK*
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:
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:
ProteinExpressed: Protein_Confirmed
SolubleProtein: Tested_Not_Soluble
Find experimental protocols in APC39020.2
Recommended expression in:
Not Applicable
Vector Name:
pMCSG19B
Synonyms:
''
Sequencing Primer:
Forward:
ANForward
Reverse:
T7 terminator
Description:
Vector for solubility of insert and for purification, with T7 promoter, adds N-terminal MBP, TVMV cleavage site, His tag, and TEV protease cleavage site; ampicillin resistance; ligation independent cloning (LIC).
Comments:
Similar to pMCSG19C except location of the TVMV CDS. See http://bioinformatics.anl.gov/mcsg/technologies/vectors.html for more details
Size (bp):
7468
Parent Vector:
None
Properties:
bacterial expression
Author Name:
Midwest Center for Structural Genomics
Midwest Center for Structural Genomics
PSI
Midwest Center for Structural Genomics
PSI
Publications:
None
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial origin
ori
pBR322 origin of replication
5432
5432
gene fragment
TVMV cds
TVMV protease coding sequence
1881
2591
promoter
T7
T7 promoter
1515
1531
protease cleavage site
TVMV
TVMV cleavage site
286
306
protease cleavage site
TEV
TEV protease cleavage site
223
243
recombination site
LIC
Ligation Independent Cloning (LIC) recombination site
204
235
repressor binding site
lac operator
LacO fragment
1490
1512
repressor binding site
tet operator
TetO fragment
2610
2765
repressor protein gene
LacI
LacI cds
3072
4031
selectable marker
AmpR
Ampicillin resistance
6220
7080
tag
His
N terminal His tag
268
285
tag
MBP
Maltose binding protein tag
337
1143
trxn termination sequence
T7 term
T7 termination sequence
26
72
