Protein coding sequences/Reporters

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Introduction to Reporters

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.

Fluorescent proteins

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 since several different fluorescent proteins are available, multiple gene expression measurements can be made in parallel.

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Name	Protein	Description	Tag	Direction	Fluorescent Color	Emission	Excitation	Length	Status	Uses
BBa_E0030	EYFP	enhanced yellow fluorescent protein derived from A. victoria GFP	None	Forward	Yellow	527	514	723	In stock	140
BBa_E0020	ECFP	engineered cyan fluorescent protein derived from A. victoria GFP	None	Forward	Cyan	476	439	723	In stock	95
BBa_E1010	mRFP1	**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)	None	Forward	Red	607	584	706	In stock	593
BBa_E0040	GFPmut3b	green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212	None	Forward	Green	511	501	720	In stock	875
BBa_E2050	mOrange	derivative of mRFP1, yeast-optimized	None		Orange	562	548	769	In stock	15
BBa_J52021		dnTraf6-linker-GFP			Green			1446	In stock	1
BBa_J52026		dnMyD88-linker-GFP			Green			1155	In stock	1
BBa_I715022		Amino Portion of RFP			Red			462	In stock	2
BBa_I715023		Carboxyl portion of RFP			Red			220	In stock	1
BBa_I712028		CherryNLS - synthetic construct monomeric red fluorescent protein with nuclear localization sequence		Forward	Red			733	In stock	8
BBa_K125500		GFP fusion brick		Forward	Green			718	In stock	13
BBa_K165005		Venus YFP, yeast optimized for fusion		Forward	Yellow			744	In stock	2
BBa_K082003	GFP	GFP(+LVA)			Green			756	In stock	31
BBa_K156009		OFP (orange fluorescent protein)		Forward	Orange			864	In stock	1
BBa_K156010		SBFP2 (strongly enhanced blue fluorescent protein)			Blue			720	In stock	9
BBa_K294055	GFPmut3b	GFP RFP Hybrid	None		Green	511	501	720	In stock	3
BBa_K283005		lpp_ompA_eGFP_streptavidin						1533	In stock	1
BBa_K157043		CFP						732	In stock	1
BBa_K411207		Green Fluorescent Protein with LVA tag (GFPLVA)			Green			753	In stock	2
BBa_K648013		GFP with Standard 25 Prefix/Suffix			Green			735	In stock	13
BBa_K525306		Fusion Protein of S-Layer SgsE and mCerulean			Blue			3129	In stock
BBa_K592100		Blue Fluorescent Protein (mTagBFP)			Blue			705	In stock	41
BBa_K592101		Yellow Fluorescent Protein (YFP)			Yellow			720	In stock	5
BBa_K864100		Super Yellow Fluorescent Protein 2 (SYFP2)			Yellow			723	In stock	8
BBa_K1122005		dsRED (BBa_E1010) compatible with Genabler assembly			Red			697	In stock
BBa_K1122006		mCherry (BBa_J06504) compatible with Genabler assembly			Red			730	In stock
BBa_K1122008		mTagBFP (BBa_K592100) compatible with Genabler assembly			Blue			721	In stock
BBa_K1033916		amajLime, yellow-green chromoprotein			Yellow, Green			693	In stock	10
BBa_K1184000		KillerRed			Red			723	In stock	5
BBa_K1159301		Super Yellow Fluorescent Protein 2 (SYFP2) in RFC[25]			Yellow			714	In stock	1
BBa_K1159302		Enhanced Cyan Fluorescent Protein (eCFP) in RFC[25]			Cyan			714	In stock	1
BBa_K1093002		sfCFP			Cyan			720	In stock
BBa_K1093003		sfBFP N-terminal			Blue			645	In stock
BBa_K1093004		sfCFP N-terminal			Cyan			645	In stock
BBa_K1093017		mCherry C-terminal			Red			243	In stock
BBa_K1399000	mRFP1	RFP from Discosoma striata (coral) with AAV-ssrA degradation tag	SsrA-AAV	Forward	Red	607	584	714	In stock
BBa_K1399001	mRFP1	RFP from Discosoma striata (coral) with LVA-ssrA degradation tag	SsrA-LVA degradation tag	Forward	Red	607	584	714	In stock	2
BBa_K1399002	mRFP1	RFP from Discosoma striata (coral) with LAA-ssrA degradation tag (wt)	SsrA-LAA degradation tag	Forward	Red	607	584	714	In stock	1
BBa_K1399003	mRFP1	RFP from Discosoma striata (coral) with DAS-ssrA degradation tag	SsrA-DAS degradation tag	Forward	Red	607	584	714	In stock	1
BBa_K1399005	GFPmut3b	GFP (mut3b) with AAV-ssrA degradation tag	SsrA-AAV degradation tag	Forward	Green	511	501	753	In stock	4
BBa_K1399004	GFPmut3b	GFP (mut3b) with LVA-ssrA degradation tag	SsrA-LVA degradation tag	Forward	Green	511	501	753	In stock	8
BBa_K1399006	GFPmut3b	GFP (mut3b) with LAA-ssrA degradation tag	SsrA-LAA degradation tag	Forward	Green	511	501	753	In stock	2
BBa_K1399007	GFPmut3b	GFP (mut3b) with SsrA-DAS+2 degradation tag	SsrA-DAS+2 degradation tag	Forward	Green	511	501	759	In stock	4
BBa_K1399008	GFPmut3b	GFP (mut3b) with DAS-ssrA degradation tag	SsrA-DAS degradation tag	Forward	Green	511	501	753	In stock	2
BBa_K1351021		Monomeric Red Fluorescent Protein from Discosoma striata			Red			675	In stock
BBa_K1323009		DsRed (RFP) Coding Sequence			Red			678	In stock	4
BBa_K1323010		YFP Coding Sequence			Yellow			720	In stock
BBa_K1510017		red florescence protein RFC23			Red			703	In stock
BBa_K1486056		CxpR & Split IFP1.4 [Cterm + Cterm][2]		Forward	Infrared			3742	In stock
BBa_K1321337		sfGFP in Freiburg format (RFC 25)			Green			711	In stock	27
BBa_K1418030		SYFP2			Yellow			723	In stock
BBa_K1418040		CFP			Cyan			723	In stock
BBa_K1418041		CFP(ns)			Cyan			717	In stock
BBa_K1740000		Dronpa 145N ORF (E. coli optimized)			Green			672	In stock
BBa_K1615089		mRFP RFC25			Red			675	In stock	11
BBa_K1742002		Dronpa 145N			Green			693	In stock	3
BBa_K1789003		GFP1(with termination codon)			Green			474	In stock	12
BBa_K1789004		GFP2			Green			249	In stock	7
BBa_K1680006		Fluorescent protein dronpa			Green			669	In stock
BBa_K1673010		Partially UV-Resistant RFP			Red			678	In stock	1
BBa_K1673101		UV-Resistant RFP			Red			678	In stock	2
BBa_K1673102		UV Mutation Optimized RFP (Improved Expression)			Red			681	In stock	1
BBa_K1673103		UV Mutation Maximizied RFP			Red			678	In stock	1
BBa_K106000		GFP, AarI BD part		Forward	Green			714	It's complicated
BBa_K106004		mCherry, Aar1 AB part			Red			708	It's complicated
BBa_K106005		mCherry, Aar1 BD part			Red			708	It's complicated
BBa_K106028		GFP, AarI AB part			Green			714	It's complicated
BBa_K125810		slr2016 signal sequence + GFP fusion for secretion of GFP		Forward	Green			779	It's complicated
BBa_K106671		GFP, Aar1 AD part			Green			714	It's complicated	7
BBa_K180008	mCherry	mCherry (rights owned by Clontech)		forward	Red			708	It's complicated	6
BBa_K1740001		Dronpa145K ORF (E. coli optimized)			Green			672	It's complicated
BBa_K1742003		Dronpa 145K			Green			741	It's complicated	3
BBa_K1819000	GFPmut3b	GFP coding sequence with an N-terminal linker		Forward	Green	511	501	750	It's complicated	1
BBa_K1602021		Green fluorescent protein (GFP) with N-terminal His-Tag			Green			738	It's complicated	2
BBa_K1602024		HisTag-GFP_Si4-Tag fusion protein			Green			807	It's complicated	2
BBa_K1857000		mVenus			Yellow			1236	It's complicated
BBa_K1680017		Cre-Dronpa Fusion			Green			1728	It's complicated
BBa_K1680018		Cre-Dronpa fusion			Green			1746	It's complicated
BBa_K1680019		Cre-dronpa fusion			Green			1764	It's complicated
BBa_K1680020		Cre-dronpa fusion			Green			1782	It's complicated
BBa_K1680021		Dronpa caged Cre with NLS			Green			2433	It's complicated
BBa_K1680022		Dronpa caged Cre with NLS			Green			2451	It's complicated
BBa_K1680023		Dronpa caged Cre with NLS			Green			2469	It's complicated
BBa_K1680024		Dronpa caged Cre with NLS			Green			2487	It's complicated
BBa_K1673104		Oxidation-Resistant RFP			Red			678	It's complicated	2
BBa_K1673410		GFP (Mutation Optimized)			Green			717	It's complicated
BBa_K2003010		Inducible Green Fluorescent Protein UnaG+6xHis-tag						462	It's complicated
BBa_K2455002	SYFP2	CPP-SYFP2	C-terminal hexa-histidine	Forward	Yellow	527	515	804	It's complicated
BBa_K2455005	mTag BFP	CPP-mTag BFP	C-terminal hexa-histidine	Forward	Blue	456	399	786	It's complicated
BBa_K2656013		sfGFP Coding Sequence						721	It's complicated	5
BBa_K2656023		GFPmut3b_LVA Coding Sequence						754	It's complicated	1
BBa_K2656014		mRFP1 Coding Sequence						679	It's complicated	3
BBa_K2656024		mRFP1_LVA Coding Sequence						715	It's complicated	1
BBa_K2656020	YFP	YFP_LVA Coding Sequence	LVA	Forward	Yellow	529	516	754	It's complicated	1
BBa_K2656021	YFP	YFP Coding Sequence	None	Forward	Yellow	529	516	721	It's complicated	1
BBa_K2558005	sfGFP	superfold GFP (+LVA_			Green			758	It's complicated	6
BBa_K3781002		mCerulean, MocloMania B3_B4						714		-1
BBa_K3781003		mVenus, MoCloMania B3_B4						714		-1
BBa_K3781015		mVenus, MocloMania B5						717		-1
BBa_K3781016		mCerulean, MocloMania B5						717		-1
BBa_K3781012		mCerulean, MocloMania B4						714		-1
BBa_K3831029		Green fluorescent protein						714		-1
BBa_K3831016		sfGFP						714		-1
BBa_K3831021		mScarlet-I						696		-1
BBa_K3831042		GFP						714		-1
BBa_K3765010		sfGFP (Superfolder GFP)						714		-1
BBa_K4139013	PbEL04	CAPE-GFP	His-Tag	Forward				912		-1
BBa_K4139015	PbEL04	COAPE - GFP	OmpA & His-Tag	Forward				984		-1
BBa_K4139017	CAP-FP	CAP-FP	His-Tag	Forward				1101		-1
BBa_K4706008	Green fluorescent GFP-like protein	eechGFP1 codon optimised S.cerevisae	none	forward			497nm	687		-1
BBa_K4759012		sfGFP						702		-1
BBa_K192001		CFP +tgt +lva						858	Not in stock
BBa_K180001	GFPmut3b	Green fluorescent protein (+LVA)	LVA	forward	Green			754	Not in stock	1
BBa_K180009	mBanana	mBanana (rights owned by Clontech)		forward	Yellow			708	Not in stock	3
BBa_K863121	GFPmut3b	green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (His-tag)	His6		Green			750	Not in stock
BBa_K863120	GFPmut3b	green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (Freiburg-Standard)	None		Green			729	Not in stock
BBa_K1491001		mTagBFP-His			Blue			723	Not in stock	1
BBa_K1491004		SuperYFP2-His			Yellow			741	Not in stock	3
BBa_K1486006		IFP[1]		Forward	Infrared			405	Not in stock	6
BBa_K1486007		IFP[2]		Forward	Infrared			585	Not in stock	6
BBa_K1362461	mRFP1	**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral) with barcode	None	Forward	Red	607	584	706	Not in stock	3
BBa_K1583017		Blue Fluorescent Protein (mTagBFP) without stopcodon			Blue			699	Not in stock	1
BBa_K1641224		mCherry-ssrA			Red			747	Not in stock	2
BBa_K1688019		dTomato, red fluorescent protein			Red			708	Not in stock	1
BBa_K1732020		GFP-HIS			Green			732	Not in stock	1
BBa_K1732021		BFPCO-His			Blue			717	Not in stock	1
BBa_K1732022		EGFPCO-His			Green			735	Not in stock	1
BBa_K1732024		YFPCO-His			Yellow			735	Not in stock	1
BBa_K1732025		RFPCO-His			Red			690	Not in stock	2
BBa_K1616016		YC155 - C terminal YFP split			Yellow			255	Not in stock	2
BBa_K1616017		YN155 - N terminal YFP split			Yellow			468	Not in stock	2
BBa_K1650038		GFP-His			Green			1159	Not in stock	1
BBa_K1799026		Discosoma RFP with stp			Red			675	Not in stock	2
BBa_K2040122		KillerRed + NLS			Red			744	Not in stock
BBa_K3128008	mRFP1	Engineered mutant of red fluorescent protein with RFC21 restriction sites	None	Forward	Red	607	584	690	Not in stock	1
BBa_K2929004	mOrange	mOrange + SpyTag	None		Orange	562	548	829	Not in stock
BBa_K3013003		mamC-FbFP BS2 fusion						819	Not in stock
BBa_K2984019		L1c-Psad-YFP-RbcS2, Expression of YFP through light-inducible promoter						1805	Not in stock
BBa_K3427000		Heat-stable fluorescent protein YFP_LOV						342	Not in stock	-1

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...

Name	Protein	Description	Direction	Uniprot	KEGG	E.C.	Substrate	Product	Length	Status
BBa_J52011		dnMyD88-linker-Rluc							1371	In stock
BBa_I712019		Firefly luciferase - luciferase from Photinus pyralis							1653	In stock
BBa_K322237		Firefly luciferase from Photinus pyralis							1653	In stock
BBa_K1159001		NanoLuc Luciferase in RFC[25]							510	In stock
BBa_K1761002		Outer Membrane Protein X (OmpX) with BsoBI-linker and NanoLuc							1241	In stock
BBa_K1689001		Coding sequence of STV-N-luc							1727	In stock
BBa_K1689006		Coding sequence of FKBP-Cluc394							875	In stock
BBa_K1680009		NanoLuc Luciferase							516	In stock
BBa_K1680010		Firefly Luciferase							1653	In stock
BBa_K1734004		Nanoluc (Codon optimized for Sf9 cells)							525	In stock
BBa_K1616025		NanoLuc							510	In stock
BBa_J52013		dnMyD88-linker-Rluc-linker-PEST191							1872	It's complicated
BBa_K525311		Fusion Protein of S-Layer SgsE and Firefly-Luciferase							4062	It's complicated
BBa_K785003		Lux(CDABEG) coding gene PoPS->Light							6427	It's complicated
BBa_K1486054		CheY/CheZ + split rLuc	Forward						3293	It's complicated
BBa_K1486055		CheY/CheZ + split fLuc	Forward						4079	It's complicated
BBa_K1732003		GLUCCO-His							522	It's complicated
BBa_K1689004		Coding sequence of Nluc398-FRB							1553	It's complicated
BBa_K1732027		GLuc-His							522	It's complicated
BBa_K2023009	Gaussia Luciferase	Gaussia luciferase coding sequence optimized for use in E.coli DH5-Alpha							561	It's complicated
BBa_K2637010		Nanoluc+PEST	Forward						642	It's complicated
BBa_K284999		Deletar este tamb�m							1431	Not in stock
BBa_K525411		Fusion Protein of S-Layer SbpA and Firefly-Luciferase							4854	Not in stock
BBa_K1761005		LargeBit Split Luciferase							477	Not in stock
BBa_K1761006		SmallBit Split Luciferase							36	Not in stock
BBa_K1732029		FireflyCO-His							1668	Not 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...

Name	Protein	Description	Direction	UniProt	KEGG	E.C.	Substrate	Product	Color	Length	Status
BBa_I732006		lacZ alpha fragment								234	In stock
BBa_I732005		lacZ (encoding beta-galactosidase, full-length)								3075	It's complicated
BBa_K147002		xylE								924	It's complicated

Miscellaneous

More...

Name	Protein	Description	Tag	Direction	UniProt	KEGG	Length	Status
BBa_K592009		amilCP, blue chromoprotein					669	In stock
BBa_K592010		amilGFP, yellow chromoprotein					699	In stock
BBa_K592011		cjBlue, green chromoprotein					702	In stock
BBa_K592012		eforRed, red chromoprotein					681	In stock
BBa_K864401		aeBlue blue chromoprotein					699	In stock
BBa_K1122007		AmilCP (BBa_K592009) compatible with Genabler assembly					685	In stock
BBa_K1033906		tsPurple, purple chromoprotein					690	In stock
BBa_K1033919		gfasPurple, purple chromoprotein					669	In stock
BBa_K1023005		eforRed					687	In stock
BBa_K1033932		spisPink, pink chromoprotein					678	In stock
BBa_K1824017		AeBlue blue chromoprotein					699	In stock
BBa_K678035		yA (green pigment) from Aspergillus nidulans					2156	It's complicated
BBa_K732001		Synthetic Chorionic Gonadotropin, beta (synCG-β)					498	It's complicated
BBa_K1824019		AeBlue blue chromoprotein with aav tag					738	It's complicated
BBa_K2656018		amilCP Coding Sequence					670	It's complicated
BBa_K4139011	Truncated PbEL04	Truncated PbEL04	Thioredoxin & His-Tag	Forward			963
BBa_K4139012	PbEL04	Full Length PbEL04	His-Tag	Forward			570
BBa_K4139018	CAPE-AFP	CAPE-AFP	His-Tag	Forward			537
BBa_K4139019	CP-MAF	Rec. CP-MAF	His-Tag	Forward			495
BBa_K4139020	Pro1	Pro1	His-Tag	Forward			1302
BBa_K5466031		AntiAFB1-scFv1 EpoR Split-N-mCerulean					1809
BBa_K5466032		AntiAFB1-scFv2 EpoR Split-C-mCerulean					1542
BBa_K5527001		amilCP purple, Purple chromoprotein					669
BBa_K5527002		amilCP yellow, Yellow chromoprotein					669
BBa_K5527003		amilCP light purple, Light purple chromoprotein					669
BBa_K1685005		aeBlue					699	Not in stock
BBa_K1685007		aeBlue with LAA tag					732	Not in stock
BBa_K1685008		aeBlue with AAV tag					732	Not in stock
BBa_K3697002		manP expression cassette					2142	Not 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> [http://openwetware.org/index.php?title=Beta-Galactosidase_Assay_%28A_better_Miller%29 Miller Assay on OpenWetWare] by Sean Moore