Difference between revisions of "Part:BBa K2797002:Design"

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===Design Notes===
 
===Design Notes===
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The <I>aad</I>A gene sequence was taken from the Addgene database (Plasmid #74703). The sequence was found to have no illegal restriction sites (i.e. no EcoRI, XbaI, SpeI, or PstI sites). A strong, standard constitutive promoter (J23119) and RBS (B0034) was added before the  <I>aad</I>A sequence. A native terminator (K2797001) was added after the <I>aad</I>A sequence to prevent any read through. The entire construct was flanked by 22 and 21 base pair biobrick prefix and suffixes and was Gibson assembled into a pSB1C3 plasmid digested with XbaI and Spe. The construct was submitted to IDT for synthesis as a gBlock.
+
The <I>aad</I>A coding sequence was taken from the Addgene database (plasmid #74703). The sequence was found to have no illegal restriction sites (i.e. no EcoRI, XbaI, SpeI, or PstI sites). A strong, standard constitutive promoter (BBa_J23119) and ribosome binding site (BBa_B0034) were added before the  <I>aad</I>A sequence. A native terminator (BBa_K2797001) was added after the <I>aad</I>A sequence. The composite part was flanked by 22 and 21 base pair BioBrick prefix and suffixes respectively. The composite part was synthesised as a gBlock by IDT. Gibson assembly was used to clone the gBlock into the pSB1C3 backbone.
  
 
===Source===
 
===Source===

Latest revision as of 23:32, 17 October 2018


Composite part for streptomycin resistance


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 861
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 709
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The aadA coding sequence was taken from the Addgene database (plasmid #74703). The sequence was found to have no illegal restriction sites (i.e. no EcoRI, XbaI, SpeI, or PstI sites). A strong, standard constitutive promoter (BBa_J23119) and ribosome binding site (BBa_B0034) were added before the aadA sequence. A native terminator (BBa_K2797001) was added after the aadA sequence. The composite part was flanked by 22 and 21 base pair BioBrick prefix and suffixes respectively. The composite part was synthesised as a gBlock by IDT. Gibson assembly was used to clone the gBlock into the pSB1C3 backbone.

Source

Addgene plasmid #74703

References

Genetic circuit design automation. Nielsen AA, Der BS, Shin J, Vaidyanathan P, Paralanov V, Strychalski EA, Ross D, Densmore D, Voigt CA. Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341. 10.1126/science.aac7341