Line 20: | Line 20: | ||
padding-left:40px; | padding-left:40px; | ||
padding-right:40px; | padding-right:40px; | ||
− | |||
} | } | ||
</style> | </style> | ||
Line 27: | Line 26: | ||
<h2>Best Part Collections</h2> | <h2>Best Part Collections</h2> | ||
− | <p style="font-size: | + | <p style="font-size: 18px;" >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 construct and test multiple variants of a given circuit. This allows teams to build, test and subsequently clone a vast number of circuit variants in a single day.</p> |
− | <p> | + | <p style="font-size: 18px;"> |
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. </p> | 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. </p> | ||
− | <p> | + | <p style="font-size: 18px;"> |
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). </p> | 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). </p> | ||
− | <p> | + | <p style="font-size: 18px;"> |
In our collection of parts, we also incorporated the ts-CI heat inducible system and TlpA heat inducible system. To use these systems, we cloned the corresponding promoter and repressor sequences needed into our standard backbone. Using these heat inducible parts, we also designed a wide variety of heat inducible circuits. Their circuits are a convenient tool for any team in the future that are using 3G methods and are pursuing this induction system.</p> | In our collection of parts, we also incorporated the ts-CI heat inducible system and TlpA heat inducible system. To use these systems, we cloned the corresponding promoter and repressor sequences needed into our standard backbone. Using these heat inducible parts, we also designed a wide variety of heat inducible circuits. Their circuits are a convenient tool for any team in the future that are using 3G methods and are pursuing this induction system.</p> | ||
− | <p> | + | <p style="font-size: 18px;"> |
While our part collection is complete for the purpose of our project, it is also open to addition. A 3G library will be an invaluable tool for other teams also hoping to implement 3G assembly, and thus the library will be open-access for all iGEM teams to contribute to. </p> | While our part collection is complete for the purpose of our project, it is also open to addition. A 3G library will be an invaluable tool for other teams also hoping to implement 3G assembly, and thus the library will be open-access for all iGEM teams to contribute to. </p> | ||
Revision as of 21:29, 15 October 2018
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 construct and test multiple variants of a given circuit. This allows teams to build, test and subsequently clone a vast number 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. To use these systems, we cloned the corresponding promoter and repressor sequences needed into our standard backbone. Using these heat inducible parts, we also designed a wide variety of heat inducible circuits. Their circuits are a convenient tool for any team in the future that are using 3G methods and are pursuing this induction system.
While our part collection is complete for the purpose of our project, it is also open to addition. A 3G library will be an invaluable tool for other teams also hoping to implement 3G assembly, and thus the library will be open-access for all iGEM teams to contribute to.
1. Promoters
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 |
2. 5' UnTranslated Regions
K2680200 | 3G BCD8 |
K2680201 | 3G BCD12 |
K2680202 | 3G BCD2 |
K2680203 | 3G B0033m |
K2680204 | 3G B0032m |
K2680205 | 3G B0034m |
3. Coding Sequences
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 |
4. Terminators
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 |
5. Basic Parts
K2680500 | WM_Pad1 |
K2680501 | WM_Pad2 |
K2680502 | Prefix BsaI Sticky End A |
K2680503 | Suffix BsaI Sticky End B |
K2680504 | Prefix BsaI Sticky End B |
K2680505 | Prefix BsaI Sticky End C (Abreviated) |
K2680506 | Prefix BsaI Sticky End C (Full) |
K2680507 | Suffix BsaI Sticky End D |
K2680508 | Prefix BsaI Sticky End D |
K2680509 | Suffix BsaI Sticky End E |
K2680510 | BsaI prefix Spacer |
K2680511 | BsaI suffix Spacer |
K2680512 | Suffix BsaI Sticky End C |
K2680513 | WM_Pad2 Construction primer |
K2680514 | WM_Pad1 Construction primer |
K2680515 | WM_Pad Insert Amplification primer forward |
K2680516 | WM_Pad Insert Amplification primer reverse |
K2680517 | WM_Pad Backbone Amplification primer forward |
K2680518 | WM_Pad Backbone Amplification primer reverse |
K2680519 | pILacO |
K2680520 | PcinAM |
K2680521 | PlasAM |
K2680522 | PluxAM |
K2680523 | PsalAM |
K2680524 | Plac/ara-1 |
K2680525 | pBAD |
K2680526 | pLsrR |
K2680527 | pLsrA |
K2680527 | pLsrA |
K2680528 | pTlpA |
K2680529 | BCD12 |
K2680530 | B0033m |
K2680531 | B0032m |
K2680532 | sfGFP |
K2680533 | L3S1P13 terminator |
K2680534 | T500 terminator |
K2680535 | ECK120033736 |
K2680536 | S. pyogenes tracrRNA terminator |