Composite

Part:BBa_K1983014:Design

Designed by: Vykintas Jaunikis   Group: iGEM16_Vilnius-Lithuania   (2016-10-13)
Revision as of 21:33, 22 October 2016 by Vykintas (Talk | contribs) (→‎Source)


PheP under constitutive promoter, high strength RBS and terminator


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 98
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was intentionally designed with XbaI site between RBS (J61132) and PheP gene (K1983013) because it was a precursor for two other parts: (K1983015) and (K1983016). These parts only differ in RBS, with (K1983014) having a high, (K1983014) having medium, and (K1983013) having low strength RBS. The two resulting parts have scars at the location of XbaI site, just as any conventional composite part.

Since this composite biobrick (K1983014) is a full construct comprised of a promoter, RBS, coding gene and terminator, with XbaI it is more adaptable regarding the regulation of this gene. Having only an XbaI site in it, it can be still be combined with other biobricks when digested with SpeI and PstI.


Primers

Primers used for amplification of the fragment:
Uni-FW: GTAGAATTCGCGGCCGCTTCTAG
Uni-RV: GTAGACTGCAGCGGCCGCTACTAG

Primers used for colony PCR screening:

For pSB1C3:
VF2: tgccacctgacgtctaagaa
VR: attaccgcctttgagtgagc


Source

This part is derived from Escherichia coli and was synthesized by Integrate DNA Technologies.

References