Difference between revisions of "Part:BBa K3110021:Design"

Latest revision as of 18:20, 21 October 2019

Weak Promoter Weak RBS lldR+lldD

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1987
Illegal AgeI site found at 619
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1424

Design Notes

Source

Weak promoter -Weak RBS-lldR flank and lldD flank-terminator were synthesized by IDT
lldR+lldD was amplified from DH5alpha genome

References

iGEM ETH zurich 2015
iGEM NUS Singapore 2016
Aguilera, L., Campos, E., Gimenez, R., Badia, J., Aguilar, J., & Baldoma, L. (2008). Dual Role of LldR in Regulation of the lldPRD Operon, Involved in L-Lactate Metabolism in Escherichia coli. Journal of Bacteriology, 190(8), 2997–3005. doi:10.1128/jb.02013-07

@@ Line 7: / Line 7: @@
 ===Design Notes===
-lldD was codon optimized to meet synthesis requirements and to remove the illegal EcoRI site present in the genomic lldD.
@@ Line 15: / Line 14: @@
 ===Source===
-lldD was Synthesized from Twist Bioscience and was joined to lldR using SOEing (Spliced Overlap Extension) PCR. This was further joined to Promoter+RBS and Terminator using SOEing (Spliced Overlap Extension) PCR.
+Weak promoter -Weak RBS-lldR flank and lldD flank-terminator were synthesized by IDT <br>
+lldR+lldD was amplified from DH5alpha genome
 ===References===
+iGEM ETH zurich 2015<br>
+iGEM NUS Singapore 2016<br>
+Aguilera, L., Campos, E., Gimenez, R., Badia, J., Aguilar, J., & Baldoma, L. (2008). Dual Role of LldR in Regulation of the lldPRD Operon, Involved in L-Lactate Metabolism in Escherichia coli. Journal of Bacteriology, 190(8), 2997–3005. doi:10.1128/jb.02013-07