Intermediate in screening plasmid v2 construction.
Assembly for J13014 by Lucy and Randy 8-05
TEST R Mobilizable Plasmid with RFP reporter = [OriTR][OnRFP]
Construction intermediate: cI857 (Lamda II) with RBS and forward terminator (B0034.J06503.B0014)
Construction intermediate: Lambda cI QPI with EYFP (Q04510.E0430)
Construction intermediate: cI857 (Lamda II) with RBS (B0034.J06503)
Abandoned in favor of using assembled cI857 QPI.
Construction intermediate: LacI ts (mut 265) QPI with Lambda cI regulated promoter (R0051.J06801)
Combines BBa_R0051 Promoter (lambda cI regulated) with BBa_J06801 QPI (LacI ts mut 265). This is an intermediate leading to BBa_J06902 and BBa_J06952.
Used with J01074 and 73 for use with testing mobilization. Test construct for mobilization. When in original cell there's constituitive cI production from a circuit on another plasmid ([R0040][J01058]). Transcription from R0051 is thus inhibited and expresssion of RFP is repressed. However, when a plasmid carrying [OriTF][r0051][StrRBS][RFP] is transferred via mobilization at OriTF, there isn't constituitive cI expression in the destination cell and thus RFP is expressed because transcription from R0051 is no longer repressed. Based off of Jaenecke et al., "A stringently controlled expression system for analysing lateral gene transfer between bacteria"
Non mobilizable TraJF, OnCFP, Key2 = [pBad/araC TraJF][DblTerminator][OnCFP][OnKey2]
Construction intermediate: Lambda cI QPI with mCherry, bact (Q04510.J06702)
Combines BBa_Q04510 QPI (Lambda cI) with BBa_J06702 Reporter (mCherry, bact). This is an intermediate leading to BBa_J06923, BBa_J06970, BBa_J06971, and BBa_J06972.
Coding region for chloramphenicol resistance
Construction intermediate: lambda cI with RBS and hybrid LacI promoter (R0011.B0034.C0051)
Constitutive lacI-production to block cI production.
Constitutive (tetR) lacI-production (lacI+LVA) to block promoter (lacI+pL) in ground state of a 2-state system. lacI+pL can be activated, e.g. over IPTG, thus leading to the production of whatever comes after it (planned: cI production as regulator in INPUT-module).
For more details see Sensing Device and Event Processing Device documentation.
TEST F mobilizable plasmid with RFP reporter = [OriTF][OnRFP]
Assembly for iGEM MIT done by Gemma and Lucy 8/17/05
TEST [OriTR][r0051][StrRBS][RFP][DblTerminator]
Test construct for mobilization. When in original cell there's constituitive cI production from a circuit on another plasmid ([R0040][J01058]). Transcription from R0051 is thus inhibited and expresssion of RFP is repressed. However, when a plasmid carrying [OriTF][r0051][StrRBS][RFP] is transferred via mobilization at OriTF, there isn't constituitive cI expression in the destination cell and thus RFP is expressed because transcription from R0051 is no longer repressed. Based off of Jaenecke et al., "A stringently controlled expression system for analysing lateral gene transfer between bacteria"
Assembly for J11009 by Lucy and Randy 8-05
Hix Left Site, recognised by Hin Recombinase to cause sequence reversal of what lies between it and the Hix Right Site.
Assembly for I1467 by Lucy and Randy 8-05
YFP under the control of the pspoIIe promoter
Construction intermediate: Lambda cI QPI with mCherry, bact (LVA+) (Q04510.J06703)
Combines BBa_Q04510 QPI (Lambda cI) with BBa_J06703 Reporter (mCherry, bact, LVA+). This is an intermediate leading to BBa_J06933, BBa_J06980, BBa_J06981, and BBa_J06982.
Construction intermediate: Lambda cI QPI with mCherry (Q04510.J06701)
Combines BBa_Q04510 QPI (Lambda cI) with BBa_J06701 Reporter (mCherry). This is an intermediate leading to BBa_J06903, BBa_J06950, BBa_J06951, and BBa_J06952.
We have some doubts about the integrity of J06701 (mCherry with RBS and T).
Assembly for J13009 by Lucy and Randy 8-05
Construction intermediate: LacI ts (mut 241) QPI with Lambda cI regulated promoter (R0051.J06800)
Combines BBa_R0051 Promoter (lambda cI regulated) with BBa_J06800 QPI (LacI ts mut 241). This is an intermediate leading to BBa_J06901 and BBa_J06951.
Construction intermediate: Lambda cI QPI with Venus YFP (Q04510.J06700)
Combines BBa_Q04510 QPI (Lambda cI) with BBa_J06700 Reporter (Venus YFP). This is an intermediate that currently does not lead to another part.
When lactose is added to the system, bind to lac repressor (endogenous) therefore GFP produced - glow green. When Tet added to the system, drive transcription of lacZ beta galactosidase which hydrolyses latose and lac repressor active. Stop Glowing green. ***NOTE*** This system will ONLY WORK IN LAC Z- bugs!!!!!
Assembly for J11019 by Lucy and Randy 8-05
Intermediate part found in Jen 1 tube 43
Terminator placed in front of I6060 (EYFP)
A terminator was placed in front of I6060 a construct containing a promoter, RBS, EYFP, and terminator. This was built as an intermediate, and is now available for use as an intermediate through the directory. The EYFP contains a LVA degradation tag.
Assembly for J13008 by Lucy and Randy 8-05
Non mobilizable TraJR, OnRFP, Key1 = [pRM TraJR][DblTerminator][OnRFP][OnKey1]
[pRM GFP] = [pRM][StrRBS][GFP][DblTerminator]
Construction intermediate: 434 cI QPI with Venus YFP (Q04520.J06700)
Combines BBa_Q04520 QPI (434 cI) with BBa_J06700 Reporter (Venus YFP). This is an intermediate that currently does not lead to another part.
Construction intermediate: 434 cI QPI with mCherry, bact (Q04520.J06702)
Combines BBa_Q04520 QPI (434 cI) with BBa_J06702 Reporter (mCherry, bact). This is an intermediate leading to BBa_J06927, BBa_J06973, BBa_J06974, and BBa_J06975.
[pRM][StrRBS][TraJR] also internally referred to as i12341
Subassembly for CalTech I12028 (S11451.S50452)
Assembly for I11008 by Lucy and Randy 8-05
Construction intermediate: 434 cI with RBS (B0034.C0052)
Assembly for part J07039 by Lucy and Randy 8-05
Mobilized OriTF with Lock1ed cI --> pRM --> GFP = [OriTF][OnLock1][cI][DblTerminator][pRM GFP]
The Hin Recombinase gene from Salmonella Typhimurium LT2. It creates Hin, which recognises Hix Left and Hix Right sites to cause sequence reversal of what lies in between.
RBS + bgl transcriptional antiterminator
Intermediate part for iGEM 7/2005 (inside J06008)
Assembly for I1467 by Randy and Lucy 8/05
It is needed in subsequent parts to observe the performance of the complete input-system, which is based on the modified lambda-system.
Construction intermediate: 434 cI QPI with mCherry (Q04520.J06701)
Combines BBa_Q04520 QPI (434 cI) with BBa_J06701 Reporter (mCherry). This is an intermediate leading to BBa_J06907, BBa_J06953, BBa_J06954, and BBa_J06955.
TEST arabinose inducible key 1 [pBad/araC][Key1]
TraJF under control of the araC pBad promoter
Construction intermediate: 434 cI with RBS and lambda cI promoter (R0051.J06570)
Construction Intermediate: mCherry, bacterial with RBS (B0034.J06504)
Combines BBa_B0034 RBS (Elowitz 1999) with BBa_J06504 mCherry, bacterial. This is an intermediate leading to BBa_J06702.
TEST arabinose inducible key 2 [pBad/araC][Key2]
Intermediate Part iGEM 7/2005 (for inside J06953)
i12351 in pSB2K3 a sacraficial kan vector used for swapping resistances by removing the onRFP indicator...loss of color as success.
TEST [pBad/araC][Key2][pTet][Lock2][RFP]
Test for activation of Lock 2 by Key 2. Need a significant increase in RFP expression for desired behavior.
Test plasmid to check for locking capability of Lock 1
Plant AtSUC2 promoter (companion cell specific)
[OriTR][OnLock??][spo0A][DblTerminator][pspoIIE YFP]
Intermediate Part (E0032.F1610) for inside parts J06007. iGEM Registry 7/2005
The part BBa_J04500 was used as a template for the creation of this new part. The -35 and -10 promoter elements were converted through mutagenesis into AflII and ClaI (respectively) restriction sites. A double restriction digest was performed and a T7 promoter was introduced in the form of primers.
We wanted a T7 promoter that could be regulated with the lac system.
The part BBa_J04500 was used as a template for the creation of this new part
Hix Right Site, recognised by Hin Recombinase to cause sequence reversal of what lies in between it and the Hix Left Site.
Coding region for tetracycline resistance
Construction Intermediate: mCherry, bacterial (LVA+) with RBS (B0034.J06505)
Combines BBa_B0034 RBS (Elowitz 1999) with BBa_J06505 mCherry, bacterial (LVA+). This is an intermediate leading to BBa_J06703.
Construction Intermediate: mCherry with forward terminator (E2060.B0015)
Combines BBa_E2060 mCherry, Ex587/Em610, yeast with BBa_B0015 Terminator (B0010, B0012). This is an intermediate leading to BBa_J06701.
Superpart BBa_J06701 does not work. This is likely to have the same problems.
Fast responding cold sensor using rapidly degrading gfp variant.
Intermediate part for iGEM 7/2005 (for inside J06017, J06018)
Construction intermediate: lambda cI with RBS (B0034.C0051)
Terminator that will eventually terminate TraJF, in front of OnCFP.
Assembly for J11008 by Lucy and Randy 8-05
Intermediate to Syncronator, Output- variants
Construction intermediate: cI857 (E07700) with RBS (B0034.J06502)
TEST [pBad/araC][Key2][pTet][Lock1][RFP]
For testing cross activation of Key 2 with Lock 1. Need to have little to no increase in RFP expression.
Strong RBS. mRFP-1 coding region. Strong Terminator.
[pspoIIe YFP] = [pspoIIe][RBS][YFP][DblTerminator]
TEST [pBAD/araC][TraJF][DblTerm][pTet][cI][DblTerm]
R0063.I0462 subassembly for I3510 and I13206
Construction intermediate: Venus YFP with forward terminator (E2030.B0015)
Combines BBa_E2030 Venus YFP, EX515/EM528, yeast with BBa_B0015 Terminator (B0010, B0012). This is an intermediate leading to BBa_J06700.
Superpart BBa_J06700 does not work. This is likely to have the same problems.
Intermediate Part for Harvard iGEM 7/2005 for inside parts J06012, J06018
Intermediate part (R0063.B0034) for inside parts J06014 and J06044
TEST [pBad/araC][StrRBS][TraJF][DblTerm][pTet][StrRBS][cI][DblTerm]
Coding region for Ampicillin resistance.
[OnLock??][spo0A][DblTerminator][pspoIIE YFP]
TEST [pBad/araC][Key1][pTet][Lock2][RFP]
For testing cross activation of Key 1 with Lock 2. Need to have little to no increase in RFP expression.
Intermediate Part for iGEM 7/2005 (for inside J06017)
TEST [pBad/araC][Key1][pTet][Lock1][RFP]
Test for activation of Lock 1 by Key 1. Need a significant increase in RFP expression for desired behavior.
Construction intermediate: 434 cI QPI with mCherry, bact (LVA+) (Q04520.J06703)
Combines BBa_Q04520 QPI (434 cI) with BBa_J06703 Reporter (mCherry, bact, LVA+). This is an intermediate leading to BBa_J06937, BBa_J06983, BBa_J06984, and BBa_J06985.
Assembly for J11005 by Lucy and Randy 8-05
Assembly for J13010 by Lucy and Randy 8-05
TEST [OriTR][pLac cI][StrRBS][RFP][DblTerm]
i12351 in pSB1AK3 a sacrificial kan vector used for swapping resistances by removing the onRFP indicator...loss of color as success.
Construction intermediate: 434 cI QPI with EYFP (Q04520.E0430)
TEST [OriTF][r0051][StrRBS][RFP][DblTerminator]
TEST [OriTF][pLam cI -][str RBS][TraJR][str RBS][GFP][Dbl Term]
TraJF prefaced by a strong ribosome binding site.
Construction intermediate: LacI QPI with Lambda cI regulated promoter (R0051.Q04121)
Combines BBa_R0051 Promoter (lambda cI regulated) with BBa_Q04121 QPI (LacI). This is an intermediate leading to BBa_J06900 and BBa_J06950.
Abandoned due to time constraints. Went ahead with only the temperature sensitive versions of the lacI QPI.
Intermediate part for iGEM 7/2005 (for inside J06950)