Status Of Building Parts

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MWSU + Davidson Status of Building Sender and Receiver

List of auto-inducers and their Sigma catalog numbers.

plux-lac hybrid promoter:

  • I751502
  • In the process of building another plux-lac hybrid promoter that is repressed by HSL and lacI

Responsible: Robert Cool, Erin Feeney and Pallavi Penumetcha

Parts we have Mini-Preps and Digestions for

Part Description Size of Insert (w/ BioBrick ends) Verification using Gel Electrophoresis Glycerol Stock
BBa_F2620 Accepts 3OC6HSL as an input and outputs PoPS 1102 bp Appears to be reversed with J04430 Yes
BBa_J04430 GFP generator switched on by IPTG 1124 bp Appears to be reversed with F2620 Yes
BBa_P0440 Produces tetR 881 bp Verified Yes
BBa_E0240 GFP generator 917 bp Verified Yes
BBa_I132623 Lux receiver that produces luxR and outputs EYFP 2085 bp Much smaller on gel Yes
BBa_I0426 Produces lasR and outputs ECFP followed by a lasR-PAI regulated promoter that outputs EYFP 2839 bp Verified Yes
BBa_C0061 Produces luxI (w/ LVA tag) 659 bp Verified Yes
BBa_C0261 Produces luxI (w/ RBS) 702 bp No successful Mini-Preps
BBa_R0062 luxR and HSL regulated promoter 96 bp Verified Yes
BBa_I0466 RhlR protein generator 983 bp Verified Yes
BBa_I0407 Produces lasI and outputs ECFP 1674 bp Verified Yes
BBa_R0079 LasR and PAI regulated promoter 198 bp Verified Yes
BBa_R0063 LuxR and HSL regulated promoter 192 bp No successful Mini-Preps
BBa_K09100 Receiver for AHL and outputs GFP 1987 bp Verified Yes
BBa_J04450 RFP coding device switched on by IPTG 2110 bp Much smaller on gel
BBa_C0079 lasR activator 797 Verified Yes
BBa_S03156 Ribosomal binding site followed by lasr coding region 840 Verified Yes
BBa_S03154 Ribosomal binding site followed by lasI coding region 726
BBa_T9002 Inputs C06HSL and outputs GFP 1986

-We will be sending all of these parts to be sequenced

Gels used to verify parts
Gel A
These are the parts run on a 0.5% gel.
Top portion of Gel A

Lane Part
1 MW
2 F0260
3 J04430
4 P0440
5 E0240
6 I13263
7 J0440

Bottom portion of Gel A

Lane Part
1 MW
2 C0061
3 I0466
4 I0407
5 I0426
6 K09100

These parts were run on a 2.5% gel.

Lane Part
1 MW
2 R0062
3 R0079
9 pTet-lac PCR product
10 MW

Look at USTC 2007 for logic gates

T9002 - no colonies

F2622 + E0240 (ligated/colonies coming up)
F2622.jpg + E0240.jpg

S03632 (ligations needed)
But BBa_S03632 is not available, so we will need to build our own version using the 3 parts below:
Parts: R0011 + C0261 + B0015
R0011b.jpg + C0261.jpg + B0015.jpg

R0011 + F1610 (minis, frag needed/colonies coming up, minis thurs)
R0011b.jpg + F1610.jpg

F2620 + E0240 (minis growing/ frag available)
F2620b.jpg + E0240.jpg