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SMa2227 (Sinorhizobium meliloti 1021) in pNIC28-Bsa4 (His-tagged bacterial expression vector)

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Explanation of Terms


Gene: Gene Symbol:  SMa2227
Gene Name:  GntR family transcriptional regulator
Sequence             
Original Clone ID: 21178
Keyword: None
Species: Sinorhizobium meliloti 1021
Type: cDNA
Vector Name: pNIC28-Bsa4               Format:  CLOSED
Source: New York Structural Genomics Research Consortium
Description: None
Comments: None
Mutation/
Discrepancy :
No / Yes
Publications: None
Authors: PSI:Biology
Albert Einstein College of Medicine
New York Structural Genomics Research Consortium
Steven Almo
SmCD00544731

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AVAILABLE
No restriction
Special MTA: PSI

 

Insert sequence: 771nts         Open reading frame : 33 to 803

TGGGTACCGAGAACCTGTACTTCCAATCCATGCCTGAGTCCGATCCGCACCTTTATGAAA
AGATCAAGGAGACCATCGACCGCCGTGTTGAAGCGGAGGAATGGCCCGCTGAGTTTCAAG
TTCCGGCGGAGGTTGAACTTGCGGAGGAGTTTGGCGCGTCACCCCTTACCGTGAGGCGTG
CATTGCGTGAACTGCAAGCCGAGGGCGTGCTCATACGCATACAAGGGCGCGGAACATTCG
TCGTTGGCCGGCGTATGCAATGTGCCGTCTTCAATATTTCCGACATGGCCGAGGAGATCG
CGCTCAGTGGCGGCGCCCATACTAGCAAGTTGATAAACCTTGGCGTAATTCCACGAGACA
GCTCGGAATCGAATATGCTGGTCCTGGGGCCGGATGGGGTCGTTTTTCATGTGCGATTGC
TGCATTTGGAGGACGGTACGCCGATCCAGCTCGAAGACCGCTATGTGAATGCAACAGAGG
CTCCGTCGTTCTTGGAGCAGGATTTCACGAAAATCACACCACATGCGTATTTGCTTCGTG
AGACGACCGTTACTTCGGTGGACAACACGATTCGCGCAATCAGGCCGGACGAAGAAGCAT
GCCGGCTTTTGCAGATCGACAGCAGCCAACCATGTCTATTGCTGGATCGATCGACCTGGC
GTGAAGGTGTTCCGGTGACACGCAGCCGCTTTATCTATCCTGGCGATCGGTACCGGCTGC
GCAGCTCACACGAGGCGCGACAGAGCCGCATCGCGACCACGCGCGGACCGGCTGCGTCTA
AGTCGCTCAAGAAAATCAAGTAACAGTAAAGGTAA


Coding Sequence Details
Insert Sequence Verified?: Y Verification Method: Sequence Verification
Mutations:
None
Discrepancy:
g319c,A107P
                                

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: Tested_Not_Found
ProteinPurified: Tested_Not_Purified
SolubleProtein: Not_Applicable
Find experimental protocols in NYCGRC-021178
Recommended expression in: DE3 RIL
       DyNA Vector Map

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Vector Name: pNIC28-Bsa4

pNIC28-Bsa4 in advanced viewed

Synonyms: pSEC-His
Sequencing Primer: Forward:  pLICf
Reverse:  pLIC rev
Description: Bacterial expression vector with T7 promoter and N-terminal 6xHis tag with TEV protease cleavage site; kanamycin resistance in bacteria; ligation independent cloning
Comments: pET expression vector with His6 tag in 22-aa N-terminal fusion peptide, with TEV protease cleavage site. Includes sites for LIC cloning, and a “stuffer” fragment that includes the SacB gene, allowing negative selection on 5% sucrose
Size (bp): 7284
Parent Vector: None
Empty Vector: None
Properties: bacterial expression, ligation independent cloning (LIC), with tag/fusion/marker
Author Name: Opher Gileadi
Publications: PMID: 20541610
Title: High-throughput production of human proteins for crystallization: the SGC experience

Vector Map:         Vector Sequence:
Sequence:

 



Vector Features:

Type Name Description Start Position End Position
bacterial origin f1 f1 origin 2266 2721
bacterial origin ColE ColE1 pBR322 origin 4338 4338
ligation independent cloning 3'LIC 2036 2050
ligation independent cloning 5'LIC 96 110
primer pLICrev pLIC reverse sequencing primer AGCAGCCAACTCAGCTTCC 2145 2164
primer pLICfor pLIC forward sequencing primer TGTGAGCGGATAACAATTCC 7262 7282
promoter T7 T7 promoter 7238 7254
protease cleavage site TEV TEV protease cleavage site 87 110
repressor binding site lac operator lac operator 7257 7281
repressor protein gene LacI Lac repressor gene 5772 6851
selectable marker KanR kanamycin resistance 2817 3629
selectable marker SacB SacB gene - allows for negative selection on 5% sucrose 569 1987
tag His N-terminal 6xHis tag 45 62
trxn termination sequence T7 term T7 terminator 2183 2229

 

  • Gene
  • Protein
  • Clone
  • Experimental
  • Organism

Gene

 

Gene Symbol : SMa2227
GENEID : 1236238
Locus Tag : SMa2227
GI : 193782752
GenBank Accession : NP_436447.2
Target GenBank: None

 

Clone

Cloning Information : 616544
Original Clone ID : 21178

Experimental

TargetTrack - Experimental data : NYCGRC-021178

Organism


TAX_ID : 266834
Species Specific ID: 1236238