Conjugation

Part:BBa_K1510915

Designed by: Kao Jung Chang   Group: iGEM14_NYMU-Taipei   (2014-10-09)

ketoisocaproate decarboxylase (THI3) Circuit

This circui encodes a branched-chain amino acid transaminase derived from the THI3 gene from Saccharomyces cerevisiae. THI3 catalyzes the second step in isoamyl alcohol biosynthesis: conversion of α-ketoisocaproate to 3-methylbutanal via enzymatic reaction.We successfully KOD PCR branched-chain amino acid transaminase(the last puzzle of part registry to build banana odor biosynthetic system) from Yeast genome.

Care picture1.jpg In the well of 4 and 6, after electrophoresis by using 1% gel, the DNA lengths are close to 2000 bp, and those are cut by using restriction enzyme EcoR1 and Spe1. The correct length of vector after cutting is 2043 bp, and the correct length of insert after cutting is 1161 bp.

Care picture2.jpg We ran taq PCR and got the result with correct DNA length (about 2000 bp), which means successfully ligation.

Care picture3.jpg This figure is a digestion check of our part. Restriction enzyme Pst1 and Xba1 is added to cut BBa_J25008 (well1~3). The result shows the correct vector lengths (3416 bp). Besides, restriction enzyme Pst1 and Xba1 is added to cut BBa_J45014(well4~8), the results shows the correct vector lengths (about 2044 bp) and insert lengths (about 1707 bp). When cut by restriction enzyme Xba1 and Pst1, the result in well 8 and 10 show that the DNA lengths are close to 2000 bp and 1500 bp. The correct length of vector after cutting is 2044 bp, and the correct length of insert after cutting is 1160 bp.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 21
    Illegal EcoRI site found at 922
    Illegal EcoRI site found at 1621
    Illegal EcoRI site found at 1807
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 21
    Illegal EcoRI site found at 922
    Illegal EcoRI site found at 1621
    Illegal EcoRI site found at 1807
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 21
    Illegal EcoRI site found at 922
    Illegal EcoRI site found at 1621
    Illegal EcoRI site found at 1807
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 21
    Illegal EcoRI site found at 922
    Illegal EcoRI site found at 1621
    Illegal EcoRI site found at 1807
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 21
    Illegal EcoRI site found at 922
    Illegal EcoRI site found at 1621
    Illegal EcoRI site found at 1807
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None