Difference between revisions of "Part:BBa K2194004:Design"

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The RBS before <i>cysPUWA</i> was designed using the Salis Lab RBS Designer [1]. The pre-sequence used was the <i>PgntK</i> promoter sequence, and the protein-coding sequence used was the <i>cysP</i> sequence. This particular RBS was chosen (see Figure 1 below) because it had the highest translation rate.
 
The RBS before <i>cysPUWA</i> was designed using the Salis Lab RBS Designer [1]. The pre-sequence used was the <i>PgntK</i> promoter sequence, and the protein-coding sequence used was the <i>cysP</i> sequence. This particular RBS was chosen (see Figure 1 below) because it had the highest translation rate.
  
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[[File:K2194004 RBS.png|600px]]
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<a href='https://static.igem.org/mediawiki/2017/1/1c/RBS_for_BBa_K2194004.png' style='color:rgb(153,117,185)'><font color='Indigo  '>BBa_K2194000</font></a>
 
<p class = "caption"> <b>Figure 1:</b> This image is taken from Salis Lab RBS Designer [1] and highlights the synthetic RBS used in BBa_K2194004.</p>
 
<p class = "caption"> <b>Figure 1:</b> This image is taken from Salis Lab RBS Designer [1] and highlights the synthetic RBS used in BBa_K2194004.</p>
  

Revision as of 22:10, 1 November 2017


Sulfate Transport System w/ Negative Feedback for Membrane Stress


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1480
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2296
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The RBS before cysPUWA was designed using the Salis Lab RBS Designer [1]. The pre-sequence used was the PgntK promoter sequence, and the protein-coding sequence used was the cysP sequence. This particular RBS was chosen (see Figure 1 below) because it had the highest translation rate.

<a href='https://static.igem.org/mediawiki/2017/1/1c/RBS_for_BBa_K2194004.png' style='color:rgb(153,117,185)'>BBa_K2194000</a>

Figure 1: This image is taken from Salis Lab RBS Designer [1] and highlights the synthetic RBS used in BBa_K2194004.

Source

See the registry pages for negative feedback membrane stress promoter PgntK (BBa_K2194002), cysPUWA proteins (BBa_K2194003), Elowitz RBS (BBa_B0034), and sulfate binding protein sbp (BBa_K2194001) sources.

The RBS before cysPUWA has the synthetic sequence 5’AAGCGGACGACACACAAGGCTCCACCAAA3’ and was designed with the “Salis Lab RBS Designer” [1].

The terminator “L3S3P21” after sbp has the synthetic sequence 5’CCAATTATTGAAGGCCTCCCTAACGGGGGGCCTTTTTTTGTTTCTGGTCTGCC3’. It is sourced from Chen et al [2] and is notable because it is a strong, recombination-resistant terminator.

References

[1] https://salislab.net/software/forward

[2] Chen, Ying-Ja et al. “Characterization of 582 Natural and Synthetic Terminators and Quantification of Their Design Constraints.” Nature Methods 10.7 (2013): 659–664. Web. 1 Nov. 2017.