Difference between revisions of "Part:BBa K3972003:Design"

 
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===Design Notes===
 
===Design Notes===
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From restriction ligation with the insert from IDT and the plasmid from Addgene pET28a-T7-ARG1
+
Insert designed and ordered from IDT. To make this part restriction and ligation reactions were performed, using the insert from IDT and the plasmid pET28a-T7-ARG1 ordered from Addgene. The ordered plasmid was not BioBrick compatible. Therefore, first mutate the sequence to make it TypeIIS BioBrick compatible. Use the following mutations: GvpC codon 25, GvpG codon 78, GvpL codon 241. Earlier mutation attempts failed.
−
 
+
−
 
+
  
 
===Source===
 
===Source===
  
−
This is a composite part including the ARG1 protein encoding region encoded in BBa_K3972002
+
This is a composite part, including the ARG1 protein and pTtrB185-269 promoter.
  
 
===References===
 
===References===

Revision as of 13:05, 20 October 2021


Tetrathionate sensor with activated pTtrB185-269-ARG1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 942
    Illegal EcoRI site found at 4095
    Illegal XbaI site found at 693
    Illegal PstI site found at 3952
    Illegal PstI site found at 4057
    Illegal PstI site found at 4435
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 942
    Illegal EcoRI site found at 4095
    Illegal NheI site found at 827
    Illegal PstI site found at 3952
    Illegal PstI site found at 4057
    Illegal PstI site found at 4435
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 942
    Illegal EcoRI site found at 4095
    Illegal BglII site found at 1446
    Illegal BamHI site found at 4851
    Illegal XhoI site found at 4539
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 942
    Illegal EcoRI site found at 4095
    Illegal XbaI site found at 693
    Illegal PstI site found at 3952
    Illegal PstI site found at 4057
    Illegal PstI site found at 4435
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 942
    Illegal EcoRI site found at 4095
    Illegal XbaI site found at 693
    Illegal PstI site found at 3952
    Illegal PstI site found at 4057
    Illegal PstI site found at 4435
    Illegal NgoMIV site found at 1897
    Illegal NgoMIV site found at 2299
    Illegal AgeI site found at 4818
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Insert designed and ordered from IDT. To make this part restriction and ligation reactions were performed, using the insert from IDT and the plasmid pET28a-T7-ARG1 ordered from Addgene. The ordered plasmid was not BioBrick compatible. Therefore, first mutate the sequence to make it TypeIIS BioBrick compatible. Use the following mutations: GvpC codon 25, GvpG codon 78, GvpL codon 241. Earlier mutation attempts failed.

Source

This is a composite part, including the ARG1 protein and pTtrB185-269 promoter.

References