Difference between revisions of "Part:BBa K1499203"
 
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<partinfo>BBa_K1499203 short</partinfo>
 
<partinfo>BBa_K1499203 short</partinfo>
  
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This part generates the serine deaminase, SdaB ([https://parts.igem.org/Part:BBa_K847080 BBa_K847080]), which confers basicity resistance with strong promoter/RBS pair ([https://parts.igem.org/Part:BBa_J23038 BBa_BBa_J23038]), terminator ([https://parts.igem.org/Part:BBa_B0010 BBa_B0010]), and the supP tRNA submitted by Stanford-Brown-Spelman 2014 ([https://parts.igem.org/Part:BBa_K1499251 BBa_K1499251]).
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This part generates the serine deaminase, SdaB ([https://parts.igem.org/Part:BBa_K847080 BBa_K847080]), which confers basicity resistance with strong promoter/RBS pair ([https://parts.igem.org/Part:BBa_J23038 BBa_BBa_J23038]), terminator ([https://parts.igem.org/Part:BBa_B0010 BBa_B0010]). However, this part will not work without the supP tRNA submitted by Stanford-Brown-Spelman 2014 ([https://parts.igem.org/Part:BBa_K1499251 BBa_K1499251]).
  
 
===Usage and Biology===
 
===Usage and Biology===
 
Serine catabolism creates buffers in the cytoplasm to help counter the effects of the high pH. This enzyme will break down serine to confer resistance to higher pH. Please refer to Stanford-Brown 2012 iGEM's page on this enzyme's part, [https://parts.igem.org/Part:BBa_K847080 BBa_K847080].  
 
Serine catabolism creates buffers in the cytoplasm to help counter the effects of the high pH. This enzyme will break down serine to confer resistance to higher pH. Please refer to Stanford-Brown 2012 iGEM's page on this enzyme's part, [https://parts.igem.org/Part:BBa_K847080 BBa_K847080].  
  
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This part is to be used in the Stanford-Brown-Spelman 2014 iGEM amberless system to confer resistance to higher pH. In wild type cells, there will be a selection against cells that express the supP tRNA, so the construct should not be expressed. The stop codons inhibit the expression of the complete product, except in amberless cells, that do not have the UAG release factors.
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This part is to be used in the Stanford-Brown-Spelman 2014 iGEM amberless system with the supP tRNA to confer resistance to higher pH. The stop codons inhibit the expression of the complete product, except in amberless cells, that do not have the UAG release factors.
  
 
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The part was sequence verified in the pSB1C3 backbone before submission to the registry. Two reads, forward and reverse, were obtained using VF2 and VR (Figure 1).
 
The part was sequence verified in the pSB1C3 backbone before submission to the registry. Two reads, forward and reverse, were obtained using VF2 and VR (Figure 1).
  
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[[Image:SBS_SdaBsequence.png|750px|thumb|center|<b>Figure 1.</b> This part was sequence verified. The silent mutations seen at the beginning of the coding region are from codon optimization that was not inputted into the reference sequence.]]
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[[Image:SBS_SdaBsequence.png|750px|thumb|center|<b>Figure 1.</b> This part was sequence verified. The silent mutations seen at the beginning of the coding region are from codon optimization that was not inputted into the reference sequence. There were three minor mutations that we do not expect to modify the function of the protein (hydrophobic to hydrophobic)]]
  
 
===Results===
 
===Results===

Latest revision as of 22:09, 2 November 2014

SdaB protein generator without tRNA + 5 amber stops

This part generates the serine deaminase, SdaB (BBa_K847080), which confers basicity resistance with strong promoter/RBS pair (BBa_BBa_J23038), terminator (BBa_B0010). However, this part will not work without the supP tRNA submitted by Stanford-Brown-Spelman 2014 (BBa_K1499251).

Usage and Biology

Serine catabolism creates buffers in the cytoplasm to help counter the effects of the high pH. This enzyme will break down serine to confer resistance to higher pH. Please refer to Stanford-Brown 2012 iGEM's page on this enzyme's part, BBa_K847080.

This part is to be used in the Stanford-Brown-Spelman 2014 iGEM amberless system with the supP tRNA to confer resistance to higher pH. The stop codons inhibit the expression of the complete product, except in amberless cells, that do not have the UAG release factors.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 6
    Illegal NheI site found at 29
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 226
    Illegal AgeI site found at 549
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 520
    Illegal SapI.rc site found at 667

Characterization

Verification of Part

The part was sequence verified in the pSB1C3 backbone before submission to the registry. Two reads, forward and reverse, were obtained using VF2 and VR (Figure 1).

Figure 1. This part was sequence verified. The silent mutations seen at the beginning of the coding region are from codon optimization that was not inputted into the reference sequence. There were three minor mutations that we do not expect to modify the function of the protein (hydrophobic to hydrophobic)

Results

This part was not investigated, but it could be verified using a basicity resistance assay.