Team:William and Mary/Part Collection

Page Title

Best Part Collections

In order to determine the appropriate expression levels of various component genes, our project required us to create and test many circuit variants. To efficiently design and clone circuits containing multiple transcriptional units, we implemented a recently designed method of circuit construction called 3G Assembly, which enables quick and modular cloning of circuits. Furthermore, mixed 3G Assembly can be used to build, test, and subsequently clone vast numbers of circuit variants in a single day. .
Since we anticipate this method to be extremely valuable to future iGEM teams, we submitted approximately 70 parts in a 3G compatible format. These parts include some of the most commonly used basic parts used by iGEM teams, as well as basic parts from our project and parts from our collaborators at UVA.
Our 3G part library consists of 4 categories: Promoter, 5’ UTR (UnTranslated Regions), Coding sequence and Terminator. Most of the variants in each category are used for tuning the relative expression level between the reporter (mScarlet) and the protease (mf-Lon). We can also easily modify degradation rate by switching the degradation tag in mScarlet, changing the strength of the promoter regulating mf-Lon, or adding a degradation tag to mf-Lon (a ssrA tag).
In our collection of parts, we also incorporated the ts-CI heat inducible system and TlpA heat inducible system, which we used extensively during the testing of our temperature sensitive IFFL circuits.
In total, while our parts collection already encompasses many of the commonly used registry parts, we hope that future teams will continue to add to it. We hope that this 3G library will prove an invaluable tool for other teams hoping to implement 3G assembly in their own labs.
3G Parts


Part ID
K2680100 3G J23103
K2680101 3G J23116
K2680102 3G J23107
K2680103 3G J23106
K2680104 3G J231026
K2680105 3G J23100
K2680106 3G pLacCIDAR
K2680107 3G pTet
K2680108 3G plLact
K2680109 3G PcinAM
K2680110 3G PlasAM
K2680111 3G PluxAM
K2680112 3G PsalAM
K2680113 3G J23115
K2680114 3G J23101
K2680115 3G pLuxR-pR
K2680116 3G pLac/ara-1
K2680117 3G pBad
K2680118 3G T7
K2680119 3G CI repressible promoter
K2680121 3G pLsrR
K2680122 3G pLsrA
K2680123 3G pTlpA
K2680124 3G pTlpAr

Coding Sequences

Part ID
K2680250 3G sfGFP
K2680251 3G mScarlet-I
K2680252 3G mScarlet-I pdt#3
K2680253 3G mScarlet-I pdt#3a
K2680254 3G mScarlet-I pdt#3b
K2680255 3G mScarlet-I pdt#3d
K2680256 3G mScarlet-I pdt#3e
K2680257 3G LacIAM
K2680258 3G LacI-ssrA
K2680259 3G TetR-ssrA
K2680260 3G eBFP2
K2680261 3G sfYFP
K2680262 3G KanR
K2680263 3G AraC-ssrA
K2680265 3G sfCFP-pdt3
K2680266 3G RFP
K2680267 3G GFP
K2680268 3G YFP
K2680269 3G cre Recombinase
K2680270 3G deCFP
K2680272 3G deGFP
K2680273 3G lambda-cI
K2680274 3G deGFP
K2680275 3G tsLambda-cI
K2680276 3G mf-Lon
K2680277 3G mf-Lon SsrA
K2680278 3G LsrR
K2680279 3G TlpA39
K2680280 3G Phage 186 integrase
K2680281 3G LsRK

5' UnTranslated Regions

Part ID
K2680200 3G BCD8
K2680201 3G BCD12
K2680202 3G BCD2
K2680203 3G B0033m
K2680204 3G B0032m
K2680205 3G B0034m


Part ID
K2680400 3G spy terminator
K2680401 3G thrL terminator
K2680402 3G L3S1P13 terminator
K2680403 3G T500_noGap, short attachment (T11)
K2680404 3G B0015
K2680405 3G ECK120033736
K2680406 3G S. pyogenes tracrRNA terminator