Intermediate part for iGEM 7/2005 (for inside J06017, J06018)
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"
Intermediate in screening plasmid v2 construction.
Construction intermediate: cI857 (Lamda II) with RBS and forward terminator (B0034.J06503.B0014)
[pRM][StrRBS][TraJR] also internally referred to as i12341
TEST [pBAD/araC][TraJF][DblTerm][pTet][cI][DblTerm]
Intermediate Part iGEM 7/2005 (for inside J06953)
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 J13009 by Lucy and Randy 8-05
Assembly for J11008 by Lucy and Randy 8-05
i12351 in pSB2K3 a sacraficial kan vector used for swapping resistances by removing the onRFP indicator...loss of color as success.
TEST R Mobilizable Plasmid with RFP reporter = [OriTR][OnRFP]
It is needed in subsequent parts to observe the performance of the complete input-system, which is based on the modified lambda-system.
For more details see Sensing Device and Event Processing Device documentation.
YFP under the control of the pspoIIe promoter
Coding region for chloramphenicol resistance
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.
TEST arabinose inducible key 2 [pBad/araC][Key2]
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
Non mobilizable TraJF, OnCFP, Key2 = [pBad/araC TraJF][DblTerminator][OnCFP][OnKey2]
Fast responding cold sensor using rapidly degrading gfp variant.
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!!!!!
Intermediate Part for iGEM 7/2005 (for inside J06017)
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: 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.
Assembly for J13010 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.
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.
i12351 in pSB1AK3 a sacrificial kan vector used for swapping resistances by removing the onRFP indicator...loss of color as success.
Assembly for J13014 by Lucy and Randy 8-05
Assembly for I1467 by Lucy and Randy 8-05
[OriTR][OnLock??][spo0A][DblTerminator][pspoIIE YFP]
Assembly for J13008 by Lucy and Randy 8-05
Mobilized OriTF with Lock1ed cI --> pRM --> GFP = [OriTF][OnLock1][cI][DblTerminator][pRM GFP]
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).
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 part J07039 by Lucy and Randy 8-05
Assembly for iGEM MIT done by Gemma and Lucy 8/17/05
Intermediate part for iGEM 7/2005 (inside J06008)
TraJF prefaced by a strong ribosome binding site.
Test plasmid to check for locking capability of Lock 1
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.
Construction intermediate: 434 cI QPI with EYFP (Q04520.E0430)
TraJF under control of the araC pBad promoter
Assembly for J11019 by Lucy and Randy 8-05
Intermediate part (R0063.B0034) for inside parts J06014 and J06044
Strong RBS. mRFP-1 coding region. Strong Terminator.
TEST F mobilizable plasmid with RFP reporter = [OriTF][OnRFP]
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.
Assembly for I11008 by Lucy and Randy 8-05
Subassembly for CalTech I12028 (S11451.S50452)
Construction intermediate: 434 cI with RBS and lambda cI promoter (R0051.J06570)
TEST arabinose inducible key 1 [pBad/araC][Key1]
Construction intermediate: cI857 (Lamda II) with RBS (B0034.J06503)
Abandoned in favor of using assembled cI857 QPI.
Terminator that will eventually terminate TraJF, in front of OnCFP.
Intermediate part for iGEM 7/2005 (for inside J06950)
Construction intermediate: lambda cI with RBS and hybrid LacI promoter (R0011.B0034.C0051)
Intermediate Part for Harvard iGEM 7/2005 for inside parts J06012, J06018
Plant AtSUC2 promoter (companion cell specific)
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.
R0063.I0462 subassembly for I3510 and I13206
[OnLock??][spo0A][DblTerminator][pspoIIE YFP]
Assembly for I1467 by Randy and Lucy 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.
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
TEST [OriTR][pLac cI][StrRBS][RFP][DblTerm]
Intermediate to Syncronator, Output- variants
Construction intermediate: 434 cI with RBS (B0034.C0052)
TEST [OriTF][r0051][StrRBS][RFP][DblTerminator]
[pspoIIe YFP] = [pspoIIe][RBS][YFP][DblTerminator]
Assembly for J11005 by Lucy and Randy 8-05
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.
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.
Construction intermediate: Lambda cI QPI with EYFP (Q04510.E0430)
[pRM GFP] = [pRM][StrRBS][GFP][DblTerminator]
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: lambda cI with RBS (B0034.C0051)
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.
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"
Assembly for J11009 by Lucy and Randy 8-05
Coding region for Ampicillin resistance.
Non mobilizable TraJR, OnRFP, Key1 = [pRM TraJR][DblTerminator][OnRFP][OnKey1]
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.
Construction intermediate: cI857 (E07700) with RBS (B0034.J06502)
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.
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.
TEST [OriTF][pLam cI -][str RBS][TraJR][str RBS][GFP][Dbl Term]
TEST [pBad/araC][StrRBS][TraJF][DblTerm][pTet][StrRBS][cI][DblTerm]
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.
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.
Intermediate part found in Jen 1 tube 43
RBS + bgl transcriptional antiterminator
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.
Intermediate Part (E0032.F1610) for inside parts J06007. iGEM Registry 7/2005
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.