Part:BBa_K4188001:Design
pSB3CY_aeBlue shuttle vector backbone for Saccharomyces cerevisiae
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 6155
Illegal XbaI site found at 3425
Illegal SpeI site found at 3790
Illegal PstI site found at 1105 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 6155
Illegal NheI site found at 3849
Illegal NheI site found at 3872
Illegal NheI site found at 4841
Illegal SpeI site found at 3790
Illegal PstI site found at 1105
Illegal NotI site found at 4782 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 6155
Illegal BamHI site found at 2441 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 6155
Illegal XbaI site found at 3425
Illegal SpeI site found at 3790
Illegal PstI site found at 1105 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 6155
Illegal XbaI site found at 3425
Illegal SpeI site found at 3790
Illegal PstI site found at 1105
Illegal AgeI site found at 793
Illegal AgeI site found at 905
Illegal AgeI site found at 2083
Illegal AgeI site found at 2406 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 1378
Illegal BsaI site found at 6161
Illegal BsaI.rc site found at 1099
Illegal SapI site found at 1082
Illegal SapI.rc site found at 6
Design Notes
During our project we faced difficulties for the transformation of our various constructs. With the given iGEM plasmids it was not possible for us to insert our gene cassettes as one of the four given places, inside the MTU, would always be occupied by a yeast specific selection marker. To circumvent this problem, we developed our own shuttle vector for yeast transformation. By inserting the selection marker on a distinct position, we were able to safe the full capacity of the MTU, further saving costs and time, by reducing the amount of needed transformations.
Source
This shuttle vector was assembled using synthesized parts, iGEM biobricks (BBa_K1430000,BBa_K864401, BBa_J23110, BBa_B0015, pSB3C01) and PCR fragments.