Status Of Building Parts

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MWSU + Davidson Status of Building Sender and Receiver

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
BBa_F2620 Accepts 3OC6HSL as an input and outputs PoPS 1102 bp Appears to be reversed with J04430
BBa_J04430 GFP generator switched on by IPTG 1124 bp Appears to be reversed with F2620
BBa_P0440 Produces tetR 881 bp Verified
BBa_E0240 GFP generator 917 bp Verified
BBa_I132623 Lux receiver that produces luxR and outputs EYFP 2085 bp Much smaller on gel
BBa_I0426 Produces lasR and outputs ECFP followed by a lasR-PAI regulated promoter that outputs EYFP 2839 bp Verified
BBa_C0061 Produces luxI (w/ LVA tag) 659 bp Verified
BBa_C0261 Produces luxI (w/ RBS) 702 bp No successful Mini-Preps
BBa_R0062 luxR and HSL regulated promoter 96 bp Verified
BBa_I0466 RhlR protein generator 983 bp Verified
BBa_I0407 Produces lasI and outputs ECFP 1674 bp Verified
BBa_R0079 LasR and PAI regulated promoter 198 bp Verified
BBa_R0063 LuxR and HSL regulated promoter 192 bp No successful Mini-Preps
BBa_K09100 Receiver for AHL and outputs GFP 1987 bp Verified
BBa_J04450 RFP coding device switched on by IPTG 2110 bp Much smaller on gel

-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.
GelA.jpg
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

GelB
These parts were run on a 2.5% gel.
GelB.jpg

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

Parts we have Glycerol Stocks for

Part Description
R0062 LuxR and HSL regulated promoter
R0063 LuxR and HSL regulate promoter
P0440 Produces tetR

Look at USTC 2007 for logic gates

T9002 - no colonies
T9002.jpg


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


S03632 (ligations needed)
S03632.jpg
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


P0440
P0440.jpg