Protein coding sequences/Reporters

< Back to Protein coding sequences

Introduction to Reporters

Cambridge 2009-RAINBOW.png

What are reporters? Why call them reporters and not fluorescent proteins, beta-galactosidase/Xgal assays or pigments? The answer is that in synthetic biology, they are often used as a mechanism to display information. If you design a biosensor system that is sensitive to substrate A, you ideally want a way of displaying these results without needing an additional assay. Using a reporter protein is a great way of achieving this property.

Reporter protein coding sequences encode proteins whose presence in the cell or organism is readily observed. For example, fluorescent proteins cause a cell to fluoresce when excited with light of a particular wavelength, luciferases cause a cell to catalyze a reaction that produces light, and enzymes such as beta-galactosidase convert a substrate to a colored product. Reporters are frequently used to quantify the strength or activity of upstream gene expression parts such as promoters and ribosome binding sites. Reporters, when fused in frame to other protein coding sequences, can also be used to identify where a protein is located in a cell or organism.

There are several different ways to measure or quantify a reporter depending on the particular reporter and what kind of characterization data is desired. Generally speaking, microscopy is useful for obtaining both spatial and temporal information on reporter activity, particularly at the single cell level. Flow cytometers are best suited for measuring the distribution in reporter activity across a large population of cells. Plate readers are generally best for taking population average measurements of many different samples over time.

Some iGEM teams have excellent projects that create new reporters, or make great use of existing ones. Please see the tables below for fluorescent proteins, chromoproteins, luciferase proteins and enzymes that produce colored substrates in the Registry.

Chromoproteins

UUChromo.jpg

Chromoproteins are often responsible for coloration in corals or sea anemones. The expressed pigment can be seen by the naked eye and without any external tool, making them very useful as reporter proteins.

Chromoproteins and fluorescent proteins can have some overlap. Generally, fluorescent proteins appear white or clear to the naked eye, but some have the property of showing pigmentation also. One such protein is BBa_E1010.

2012 Uppsala University: The Uppsala University team created a chromoprotein collection as part of their project in 2012. Here are pellets of bacteria expressing chromoproteins eforRed (BBa_K592012), RFP (BBa_E1010), cjBlue (BBa_K592011), aeBlue (BBa_K864401), amilGFP (BBa_K592010) and amilCP (BBa_K592009).


To add parts to this table use the category: //cds/reporter/chromoprotein


More...
NameDescriptionLengthCreated byDocumentationUsesTypeStatus
BBa_K592009amilCP, blue chromoprotein669Lei Sun222331CodingIn stock
BBa_K592011cjBlue, green chromoprotein702Antonio Ascue Avalos20856CodingIn stock
BBa_K1122007AmilCP (BBa_K592009) compatible with Genabler assembly685Lina Gasiūnaitė1132 CodingIn stock
BBa_K1033902meffBlue, blue chromoprotein669Erik Gullberg18851CodingIn stock
BBa_K1033906tsPurple, purple chromoprotein690Sabri Jamal 11331CodingIn stock
BBa_K1033916amajLime, yellow-green chromoprotein693Sabri Jamal 20876CodingIn stock
BBa_K1033910fwYellow, yellow chromoprotein714Sabri Jamal13382CodingIn stock
BBa_K1023005eforRed687Michiel Karrenbelt2758 CodingIn stock
BBa_K1399002RFP from Discosoma striata (coral) with LAA-ssrA degradation tag (wt)714Anna Stikane  CodingIn stock
BBa_K1399000RFP from Discosoma striata (coral) with AAV-ssrA degradation tag714Anna Stikane  CodingIn stock
BBa_K1399003RFP from Discosoma striata (coral) with DAS-ssrA degradation tag714Anna Stikane  CodingIn stock
BBa_K1399001RFP from Discosoma striata (coral) with LVA-ssrA degradation tag714Anna Stikane 1CodingIn stock
BBa_K592010amilGFP, yellow chromoprotein699Lei Sun196211CodingIt's complicated
BBa_K592012eforRed, red chromoprotein 681Lei Sun18857CodingIt's complicated
BBa_K864401aeBlue blue chromoprotein699Erik Lundin22166CodingIt's complicated
BBa_K1824017AeBlue blue chromoprotein699Zhe Yang  Coding 
BBa_K1824019AeBlue blue chromoprotein with aav tag738Zhe Yang  Coding 
BBa_K1685005aeBlue699ITB_Indonesia 2015 2Coding 
BBa_K1680006Fluorescent protein dronpa669Katharina Sporbeck  Coding 
BBa_K1680017Cre-Dronpa Fusion1728Nicolai von Kügelgen  Coding 
BBa_K1680018Cre-Dronpa fusion1746Nicolai von Kügelgen  Coding 
BBa_K1680019Cre-dronpa fusion1764Nicolai von Kuegelgen  Coding 
BBa_K1680020Cre-dronpa fusion1782Nicolai von Kuegelgen  Coding 
BBa_K1680021Dronpa caged Cre with NLS2433Nicolai von Kuegelgen  Coding 
BBa_K1680022Dronpa caged Cre with NLS2451Nicolai von Kuegelgen  Coding 
BBa_K1680023Dronpa caged Cre with NLS2469Nicolai von Kuegelgen  Coding 
BBa_K1680024Dronpa caged Cre with NLS2487Nicolai von Kuegelgen  Coding 
BBa_K1886004sll1472 ccaS2733Lejian Jiang -1Coding 
BBa_K1033919gfasPurple, purple chromoprotein669Sabri Jamal 16243CodingNot in stock
BBa_K1033932spisPink, pink chromoprotein678Erik Lundin2013 CodingNot in stock
BBa_K1033933asPink, pink chromoprotein702Erik Lundin22911CodingNot in stock
BBa_K1484000AsPink chromoprotein in GoldenGate standard721Virginie Rutten  CodingNot in stock
BBa_K1484001tsPurple chromoprotein in GoldenGate standard709Virginie Rutten  CodingNot in stock
BBa_K1484002AmajLime chromoprotein in BBa and MoClo standard711Virginia Rutten  CodingNot in stock
BBa_K1484003AeBlue chromoprotein in BBa and MoClo standard724----  CodingNot in stock
BBa_K1484004AmilGFP chromoprotein in BBa and MoClo standard724Virginia Rutten  CodingNot in stock
BBa_K1484005AmilCP chromoprotein in BBa and MoClo standard706Virginia Rutten  CodingNot in stock
BBa_K1484006EforRed chromoprotein in BBa and MoClo standard706Virginia Rutten  CodingNot in stock
BBa_K1484007cjBlue chromoprotein in BBa and MoClo standard724Virginia Rutten  CodingNot in stock
BBa_K1484008AeBlue0034 chromoprotein in BBa and MoClo standard758Virginia Rutten  CodingNot in stock
BBa_K1362461**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral) with barcode 706Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha 3CodingNot in stock


IP-Free Fluorescent proteins

DNA2.0 Four protein Screen Shot 2014-02-05 at 4.57.24 PM.png

These parts were created by DNA 2.0 as part of their IP-Free series of fluorescent and chromogenic proteins. They are available to use under the BioBrick Public Agreement.

To add parts to this table, make sure to add the category: //legal/ip-free.


More...
NameDescriptionLengthCreated byDocumentationUsesTypeStatus
BBa_J97003IP-Free TannenRFP (Red Fluorescent Protein)678Drew Endy5625 ReporterNot in stock
BBa_J97002IP-Free BlazeYFP (Yellow Fluorescent Protein)708Drew Endy5628 ReporterNot in stock
BBa_J97000IP-Free EiraCFP (Cyan Fluorescent Protein)699Drew Endy5681 ReporterNot in stock
BBa_J97001P-Free JuniperGFP (Green Fluorescent Protein)702Drew Endy5644 ReporterNot in stock

Fluorescent proteins

GFP visualized under UV light.
Fluorescent proteins are convenient ways to visualize or quantify the output of a device or part. Many different fluorescent proteins are available from the Registry. The first fluorescent protein to be cloned and the most commonly used fluorescent protein is green fluorescent protein (GFP). GFP was cloned from jellyfish. Based on GFP, several investigators generated mutations that altered the spectral properties of GFP resulting in yellow fluorescent protein (YFP) and cyan fluorescent protein (CFP).
RFP visualized in white light.
Red fluorescent protein (RFP), shown on the left, was isolated from coral. Note that the colonies appear red to the naked eye as well as fluoresce red. RFP is unrelated to the GFP family of fluorescent proteins. Several derivatives of RFP have been generated including mOrange and mCherry.

Fluorescence can be readily quantified using a microscope, plate reader or flow cytometer equipped to excite the fluorescent protein with the appropriate wavelength of light. Since several different fluorescent proteins are available, multiple gene expression measurements can be made in parallel.

To include a part in this table, include the categories "//function/reporter/fluorescence" and set the part type to Coding under the Hard Information tab of the part.


More...
NameProteinDescriptionTagDirectionFluorescent ColorEmissionExcitationLengthStatus
BBa_E0030EYFPenhanced yellow fluorescent protein derived from A. victoria GFPNoneForwardYellow527514723In stock
BBa_E0020ECFPengineered cyan fluorescent protein derived from A. victoria GFP NoneForwardCyan476439723In stock
BBa_E1010mRFP1 **highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)NoneForwardRed607584706In stock
BBa_E2050mOrangederivative of mRFP1, yeast-optimizedNoneOrange562548769In stock
BBa_E0040GFPmut3b green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212NoneForwardGreen511501720In stock
BBa_J52021 dnTraf6-linker-GFPGreen  1446In stock
BBa_J52026 dnMyD88-linker-GFPGreen  1155In stock
BBa_I715022 Amino Portion of RFPRed  462In stock
BBa_I715023 Carboxyl portion of RFPRed  220In stock
BBa_K125500 GFP fusion brickForwardGreen  718In stock
BBa_K165005 Venus YFP, yeast optimized for fusionForwardYellow  744In stock
BBa_K082003GFPGFP(+LVA)Green  756In stock
BBa_K294055GFPmut3bGFP RFP HybridNoneGreen511501720In stock
BBa_K411207 Green Fluorescent Protein with LVA tag (GFPLVA)Green  753In stock
BBa_K648013 GFP with Standard 25 Prefix/Suffix Green  735In stock
BBa_K525306 Fusion Protein of S-Layer SgsE and mCeruleanBlue  3129In stock
BBa_K592100 Blue Fluorescent Protein (mTagBFP)Blue  705In stock
BBa_K592101 Yellow Fluorescent Protein (YFP)Yellow  720In stock
BBa_K864100 Super Yellow Fluorescent Protein 2 (SYFP2)Yellow  723In stock
BBa_K1122005 dsRED (BBa_E1010) compatible with Genabler assemblyRed  697In stock
BBa_K1122006 mCherry (BBa_J06504) compatible with Genabler assemblyRed  730In stock
BBa_K1122008 mTagBFP (BBa_K592100) compatible with Genabler assemblyBlue  721In stock
BBa_K1033916 amajLime, yellow-green chromoproteinYellow, Green  693In stock
BBa_K1184000 KillerRedRed  723In stock
BBa_K1159301 Super Yellow Fluorescent Protein 2 (SYFP2) in RFC[25]Yellow  714In stock
BBa_K1159302 Enhanced Cyan Fluorescent Protein (eCFP) in RFC[25]Cyan  714In stock
BBa_K1093002 sfCFPCyan  720In stock
BBa_K1093003 sfBFP N-terminalBlue  645In stock
BBa_K1093004 sfCFP N-terminalCyan  645In stock
BBa_K1093017 mCherry C-terminalRed  243In stock
BBa_K1399002mRFP1RFP from Discosoma striata (coral) with LAA-ssrA degradation tag (wt)SsrA-LAA degradation tagForwardRed607584714In stock
BBa_K1399000mRFP1RFP from Discosoma striata (coral) with AAV-ssrA degradation tagSsrA-AAVForwardRed607584714In stock
BBa_K1399003mRFP1RFP from Discosoma striata (coral) with DAS-ssrA degradation tagSsrA-DAS degradation tagForwardRed607584714In stock
BBa_K1399005GFPmut3bGFP (mut3b) with AAV-ssrA degradation tagSsrA-AAV degradation tagForwardGreen511501753In stock
BBa_K1399001mRFP1RFP from Discosoma striata (coral) with LVA-ssrA degradation tagSsrA-LVA degradation tagForwardRed607584714In stock
BBa_K1399004GFPmut3bGFP (mut3b) with LVA-ssrA degradation tagSsrA-LVA degradation tagForwardGreen511501753In stock
BBa_K1399006GFPmut3bGFP (mut3b) with LAA-ssrA degradation tagSsrA-LAA degradation tagForwardGreen511501753In stock
BBa_K1399007GFPmut3bGFP (mut3b) with SsrA-DAS+2 degradation tagSsrA-DAS+2 degradation tagForwardGreen511501759In stock
BBa_K1399008GFPmut3bGFP (mut3b) with DAS-ssrA degradation tagSsrA-DAS degradation tagForwardGreen511501753In stock
BBa_K1351021 Monomeric Red Fluorescent Protein from Discosoma striataRed  675In stock
BBa_K1323009 DsRed (RFP) Coding SequenceRed  678In stock
BBa_K1323010 YFP Coding SequenceYellow  720In stock
BBa_K1510017 red florescence protein RFC23Red  703In stock
BBa_K1486056 CxpR & Split IFP1.4 [Cterm + Cterm][2]ForwardInfrared  3742In stock
BBa_K1321337 sfGFP in Freiburg format (RFC 25) Green  711In stock
BBa_K1418030 SYFP2Yellow  723In stock
BBa_K1418040 CFPCyan  723In stock
BBa_K1418041 CFP(ns)Cyan  717In stock
BBa_I712028 CherryNLS - synthetic construct monomeric red fluorescent protein with nuclear localization sequenceForwardRed  733It's complicated
BBa_K106000 GFP, AarI BD partForwardGreen  714It's complicated
BBa_K106004 mCherry, Aar1 AB partRed  708It's complicated
BBa_K106005 mCherry, Aar1 BD partRed  708It's complicated
BBa_K106028 GFP, AarI AB partGreen  714It's complicated
BBa_K125810 slr2016 signal sequence + GFP fusion for secretion of GFPForwardGreen  779It's complicated
BBa_K156009 OFP (orange fluorescent protein)ForwardOrange  864It's complicated
BBa_K156010 SBFP2 (strongly enhanced blue fluorescent protein)Blue  720It's complicated
BBa_K106671 GFP, Aar1 AD partGreen  714It's complicated
BBa_K180008mCherrymCherry (rights owned by Clontech)forwardRed  708It's complicated
BBa_K1740000 Dronpa 145N ORF (E. coli optimized)Green  672 
BBa_K1740001 Dronpa145K ORF (E. coli optimized) Green  672 
BBa_K1615089 mRFP RFC25Red  675 
BBa_K1742002 Dronpa 145NGreen  693 
BBa_K1742003 Dronpa 145KGreen  741 
BBa_K1819000GFPmut3bGFP coding sequence with an N-terminal linkerForwardGreen511501750 
BBa_K1583017 Blue Fluorescent Protein (mTagBFP) without stopcodonBlue  699 
BBa_K1789003 GFP1(with termination codon)Green  474 
BBa_K1789004 GFP2 Green  249 
BBa_K1641224 mCherry-ssrARed  747 
BBa_K1688019 dTomato, red fluorescent proteinRed  708 
BBa_K1602021 Green fluorescent protein (GFP) with N-terminal His-TagGreen  738 
BBa_K1602024 HisTag-GFP_Si4-Tag fusion proteinGreen  807 
BBa_K1857000 mVenusYellow  1236 
BBa_K1732020 GFP-HISGreen  732 
BBa_K1680006 Fluorescent protein dronpaGreen  669 
BBa_K1680017 Cre-Dronpa FusionGreen  1728 
BBa_K1680018 Cre-Dronpa fusionGreen  1746 
BBa_K1680019 Cre-dronpa fusionGreen  1764 
BBa_K1680020 Cre-dronpa fusionGreen  1782 
BBa_K1680021 Dronpa caged Cre with NLSGreen  2433 
BBa_K1680022 Dronpa caged Cre with NLSGreen  2451 
BBa_K1680023 Dronpa caged Cre with NLSGreen  2469 
BBa_K1680024 Dronpa caged Cre with NLSGreen  2487 
BBa_K1732021 BFPCO-HisBlue  717 
BBa_K1673010 Partially UV-Resistant RFPRed  678 
BBa_K1673101 UV-Resistant RFPRed  678 
BBa_K1673102 UV Mutation Optimized RFP (Improved Expression)Red  681 
BBa_K1673103 UV Mutation Maximizied RFPRed  678 
BBa_K1673104 Oxidation-Resistant RFPRed  678 
BBa_K1732022 EGFPCO-HisGreen  735 
BBa_K1673410 GFP (Mutation Optimized)Green  717 
BBa_K1732025 RFPCO-HisRed  690 
BBa_K1732024 YFPCO-HisYellow  735 
BBa_K1616016 YC155 - C terminal YFP splitYellow  255 
BBa_K1616017 YN155 - N terminal YFP splitYellow  468 
BBa_K1650038 GFP-HisGreen  1159 
BBa_K1799026 Discosoma RFP with stpRed  675 
BBa_K2003010 Inducible Green Fluorescent Protein UnaG+6xHis-tag  462 
BBa_K2040122 KillerRed + NLSRed  744 
BBa_K1486006 IFP[1]ForwardInfrared  405Not in stock
BBa_K192001 CFP +tgt +lva  858Not in stock
BBa_K180001GFPmut3bGreen fluorescent protein (+LVA)LVAforwardGreen  754Not in stock
BBa_K283005 lpp_ompA_eGFP_streptavidin  1533Not in stock
BBa_K180009mBananamBanana (rights owned by Clontech)forwardYellow  708Not in stock
BBa_K157043 CFP  732Not in stock
BBa_K863121GFPmut3bgreen fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (His-tag)His6Green  750Not in stock
BBa_K863120GFPmut3bgreen fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (Freiburg-Standard)NoneGreen  729Not in stock
BBa_K1491004 SuperYFP2-HisYellow  741Not in stock
BBa_K1491001 mTagBFP-HisBlue  723Not in stock
BBa_K1486007 IFP[2]ForwardInfrared  585Not in stock
BBa_K1362461mRFP1**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral) with barcode NoneForwardRed607584706Not in stock


Luciferases

Luminescence can be readily quantified using a plate reader or luminescence counter. Luciferases are especially well-suited for measuring low levels of gene expression because cells tend to have little to no background luminescence in the absence of a luciferase.


More...
NameProteinDescriptionDirectionUniprotKEGGE.C.SubstrateProductLengthStatus
BBa_J52011 dnMyD88-linker-Rluc     1371In stock
BBa_I712019 Firefly luciferase - luciferase from Photinus pyralis     1653In stock
BBa_K322237 Firefly luciferase from Photinus pyralis     1653In stock
BBa_K1159001 NanoLuc Luciferase in RFC[25]     510In stock
BBa_J52013 dnMyD88-linker-Rluc-linker-PEST191     1872It's complicated
BBa_K525311 Fusion Protein of S-Layer SgsE and Firefly-Luciferase     4062It's complicated
BBa_K785003 Lux(CDABEG) coding gene PoPS->Light     6427It's complicated
BBa_K1486054 CheY/CheZ + split rLucForward     3293It's complicated
BBa_K1486055 CheY/CheZ + split fLucForward     4079It's complicated
BBa_K1689001 Coding sequence of STV-N-luc     1727 
BBa_K1732003 GLUCCO-His     522 
BBa_K1761002 Outer Membrane Protein X (OmpX) with BsoBI-linker and NanoLuc     1241 
BBa_K1761005 LargeBit Split Luciferase       
BBa_K1689004 Coding sequence of Nluc398-FRB      1553 
BBa_K1689006 Coding sequence of FKBP-Cluc394     875 
BBa_K1680009 NanoLuc Luciferase     516 
BBa_K1680010 Firefly Luciferase     1653 
BBa_K1761006 SmallBit Split Luciferase       
BBa_K1732029 FireflyCO-His     1668 
BBa_K1734004 Nanoluc (Codon optimized for Sf9 cells)     525 
BBa_K1616025 NanoLuc      510 
BBa_K2023009Gaussia LuciferaseGaussia luciferase coding sequence optimized for use in E.coli DH5-Alpha     561 
BBa_K284999 Deletar este também     1431Not in stock
BBa_K525411 Fusion Protein of S-Layer SbpA and Firefly-Luciferase     4854Not in stock
BBa_K1732027 GLuc-His     522Not in stock


Enzymes that produce colored substrates

Enzymes that produce colored substrates can be quantified using spectrophotometers or other instruments that can take absorbance measurements including plate readers. Like luciferases, enzymes like β-galactosidase are good for measuring low levels of gene expression because they tend to amplify low signals.


More...
NameProteinDescriptionDirectionUniProtKEGGE.C.SubstrateProductColorLengthStatus
BBa_I732006 lacZ alpha fragment     234In stock
BBa_I732005 lacZ (encoding beta-galactosidase, full-length)     3075It's complicated
BBa_K147002 xylE     924It's complicated


Miscellaneous


More...
NameProteinDescriptionTagDirectionUniProtKEGGLengthStatus
BBa_K592009 amilCP, blue chromoprotein  669In stock
BBa_K592011 cjBlue, green chromoprotein  702In stock
BBa_K1122007 AmilCP (BBa_K592009) compatible with Genabler assembly  685In stock
BBa_K1033906 tsPurple, purple chromoprotein  690In stock
BBa_K1023005 eforRed  687In stock
BBa_K678035 yA (green pigment) from Aspergillus nidulans  2156It's complicated
BBa_K592010 amilGFP, yellow chromoprotein  699It's complicated
BBa_K592012 eforRed, red chromoprotein   681It's complicated
BBa_K864401 aeBlue blue chromoprotein  699It's complicated
BBa_K732001 Synthetic Chorionic Gonadotropin, beta (synCG-β)  498It's complicated
BBa_K1685007 aeBlue with LAA tag  732 
BBa_K1685008 aeBlue with AAV tag  732 
BBa_K1824017 AeBlue blue chromoprotein  699 
BBa_K1824019 AeBlue blue chromoprotein with aav tag  738 
BBa_K1685005 aeBlue  699 
BBa_K1033919 gfasPurple, purple chromoprotein  669Not in stock
BBa_K1033932 spisPink, pink chromoprotein  678Not in stock

References

<biblio>

  1. Pardy pmid=7866866

</biblio>

Fluorescent proteins

<biblio>

  1. Chalfie pmid=8303295
  2. Prasher pmid=1347277
  3. Cormack pmid=8707053
  4. Tsien pmid=9759496
  5. Andersen pmid=9603842
  6. Leveau pmid=11698362
  7. Iafolla pmid=18350571
  8. Dong pmid=18352052
  9. Bagh pmid=18352063
  10. Baird pmid=11050229
  11. Patterson pmid=15583657

</biblio>

Luciferase

<biblio>

  1. Bronstein pmid=8080073

</biblio>

β-galactosidase

<biblio>

  1. Griffith pmid=11779182
  2. Villarejo pmid=4552692
  3. Zamenhof pmid=4552986
  4. Ullman pmid=1345751

</biblio> Miller Assay on OpenWetWare by Sean Moore