Cloning & Expression

MAT System Vectors


The new MAT tag or Metal Affinity Tag (HNHRHKH) provides for highly sensitive detection using the Anti-MAT™ antibody and robust one-step purification of recombinant MAT fusion proteins using HIS-Select™ Nickel and Cobalt Affinity Gels.

 

 

Bacterial Expression-tac Promoter


  • Cytoplasmic expression
  • N- or C- terminal fusion proteins
  • Ampicillin resistance, T1T2 transcriptional terminator, pMB1(derivative of pBR322) origin of replication, and lacI gene for repression of the tac promoter
  • Expression levels using the tac promoter may be in excess of 10 mg/L of culture when using IPTG as a de-repressor
  • Expression in any established Escherichia coli expression host
 
Product MCS Region
E5530, pTAC-MAT-1
N-terminal MAT
 
E5405, pTAC-MAT-2
C-terminal MAT
 

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Bacterial Expression-T7 Promoter


  • Cytoplasmic expression
  • N- or C- terminal fusions, dual tag configurations
  • FLAG® vectors for highly sensitive detection and purification using ANTI-FLAG® antibodies, resins, and plates
  • Ampicillin resistance, T1T2 transcriptional terminator, pMB1 (derivative of pBR322) origin of replication, and lacI gene for repression of the T7 promoter
  • Higher yields of recombinant protein than tac system
  • lac operator (lacO) sequences reduce leaky expression
  • Requires hosts with a source of the T7 polymerase such as (DE3) lysogenic strains
 

Product MCS Region
E5780, pT7-MAT™-1
N-terminal MAT

E5655, pT7-MAT™-2
C-terminal MAT
 
E5280, pT7-FLAG-MAT™-1
N-terminal Met-FLAG, C-terminal MAT (dual tag)

E4905, pT7-MAT™-FLAG-2
N-terminal MAT,C-terminal FLAG (dual tag)

E5155, pT7-MAT-FLAG™-1
N-terminal MAT-FLAG (dual tag)

E5030, pT7-FLAG-MAT™-2
C-terminal FLAG-MAT (dual tag)


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Mammalian Expression


  • Transient, cytoplasmic expression
  • N- or C- terminal dual tag configurations
  • FLAG® vectors for highly sensitive detection and purification using ANTI-FLAG® antibodies, resins, and plates
  • Strong human cytomegalovirus (CMV) promoter regulatory region drives constitutive protein expression levels as high as 1 mg/L in COS cells (in less potent cell lines, protein levels are typically ~0.1 mg/L)
  • pMB1 (derivative of pBR322) origin for replication in bacterial cells
  • Ampicillin resistance selection in bacteria
 
Product MCS Region
C5864,
N-terminal Met-FLAG, C-terminal MAT (Dual tagged)
 
C5989, pCMV-MAT-FLAG™-1
N-terminal MAT-FLAG (Dual tagged)

C6114, pCMV-FLAG-MAT™-2
C-terminal FLAG-MAT (Dual tagged)

 

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Insect Cell Expression


  • N-terminal and C-terminal configurations
  • Compatible with commercially available ORF 1629 deleted baculoviral DNA
  • Strong polyhedrin (polh) promoter results in high-level expression of target genes during the very late phase of infection
  • AcNPV ORF 603 and ORF1629 regions flank the polh promoter and MCS for homologous recombination with baculoviral DNA and generation of recombinant baculovirus
  • pUC origin of replication and β-lactamase gene (ampr for propagation and selection in bacterial cells
 
Product MCS Region
T6699, pPolh-MAT™-1
N-terminal MAT

T6574, pPolh-MAT™-1
C-terminal MAT
 

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