Difference between revisions of "Team:William and Mary/Part Collection"

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<h2>Best Part Collections</h2>
 
<h2>Best Part Collections</h2>
  
<p style="font-size: 5px;">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 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

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 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

Name
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

2. 5' UnTranslated Regions

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

3. Coding Sequences

Name
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

4. Terminators

Name
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

5. Basic Parts

Name
Part ID
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