Coding

Part:BBa_K1431101:Design

Designed by: Rifei Chen, Yushan Zhang   Group: iGEM14_SUSTC-Shenzhen   (2014-10-09)
Revision as of 03:25, 18 October 2014 by ChenRifei (Talk | contribs) (Design Notes)

TetOn-3G, an ideal controller of mammalian gene expression with TRE-3G promoter+PolyA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Design the primers and PCR by Q5® High-Fidelity DNA Polymerases from plasmid.

Plasmid_transfected.jpg
pBX-093 PB5-HS4-SV40-puro-2A-tetON3G-pA-HS4-TRE-AzaminGreen-2A-T

Primer Forward: T TCTAGA TGGGATCAAGACTGGACAAGA
Primer Reverse: AAAACTGCAG CGGCCGC T ACTAGT A CCGAAGCCCAACCTTTCATA

Sequencing Results

We sent fresh bacteria broth for sequencing using standard Biobricks sequencing primer VF2/VR. The sequencing cooperation we used is Invitrogen Guangzhou filiale.

Sequence of sequencing primer we used:
VF2: tgccacctgacgtctaagaa
VR: attaccgcctttgagtgagc

The result shows the same sequence with our ideal design.

Source

The plasmid was from our instructor, Huangwei's lab. And we design primers to copy down by PCR.

References