Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila.

Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
Cell Rep. 2021 Sep 14;36(11):109700. doi: 10.1016/j.celrep.2021.109700. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
165773	p3xP3-Hsp70b	HSP70B Promoter fused to synthetic 3xP3 eye driver
165774	pMCS-TCCC	Multiple cloning site part for enhancer cloning, requires minimal promoter element
165775	5xGMR	5xGMR syntethic eye driver enhancer, requires minimal promoter element
165776	pdSCP	Minimal synthetic Drosophila core promoter, requires enhancer element for full expression
165777	pDmActin5c	Drosophila Actin 5C promoter
165779	p3xP3	3xP3 eye driver enhancer element, requires promoter element for function
165780	pDmHsp70b-TACT	HSP70B promoter for fusion with bacterial promoters
165781	pDmAlfaTub84b	Drosophila Alpha Tubulin promoter
165783	pDmVasa	Drosohphila vasa germline promoter and 5'UTR
165785	pDmHsp70b	Drosophila HSP70B promoter fragment, shows basal expression that can be further enhanced via heatshock
165786	pDmPolyUbiq	Drosophila Polyubiquitin promoter, strongest promoter in the library
165787	Promoterless TP10	Drosphila TP10 poly(a) terminator sequence in promoter grammar, for assembly of no expression control vectors
165790	pPromoterless-P2A	Viral P2A part in promoter grammar, for building promoterless control vectors
165791	pR20A02	GB2.0 compatible FlyLight Enhancer element R20A02 in enhancer grammar
165792	pR70B04	GB2.0 compatible FlyLight Enhancer element R70B04 in enhancer grammar
165793	pR76H03	GB2.0 compatible FlyLight Enhancer element R76H03in enhancer grammar
165794	pCP6-Dros	Fusion grammar bacterial CP6 promoter, assembles with pHSP70B-TACT
165795	pCP6-STD	Standard grammar bacterial CP6 promoter
165796	pCP7-Dros	Fusion grammar bacterial CP7 promoter, assembles with pHSP70B-TACT
165797	pTP10	Drosophila TP10 poly(a) terminator element
165798	pSV40L	Viral Simian virus 40 Late poly(a) terminator element
165800	pAdh 3'UTR Short	Drosophila HSP70B 3'UTR sequence
165801	pHSV-TK	Minimal viral terminator derived from Herpes Simplex 1 thymidine kinase gene
165803	prrnBT1_STD	Bacterial rrnB gene T1 terminator
165804	prrnBT2_STD	Bacterial rrnB gene T2 terminator
165805	pL3S2P21_STD	Synthetic bacterial terminator
165806	pspy_STD	bacterial terminator
165807	pGGGSx4	Flexible peptide linker of 4 repeats of GGGS, for assembly of TL1 to TL5 parts
165809	pT2A	Viral T2A linker element for assembly of TL3 to TL5 parts
165810	pGGSx4 NT	Flexible peptide linker of 4 repeats of GGS, for assembly of N-terminal TL1 tags to TL3 parts
165811	pGGSx4 CT	Flexible peptide linker of 4 repeats of GGS, for assembly of C-terminal TL5 tags to TL4 parts
165814	p4xLexAOp-A1	Alpha1 4xLexAOp for assembly of larger LexAOp response elements
165815	p4xLexAOp-A2	Alpha2 4xLexAOp for assembly of larger LexAOp response elements
165816	p4xLexAOp-O2	Omega2 4xLexAOp for assembly of larger LexAOp response elements
165817	p4xLexAOp-O1	Omega1 4xLexAOp for assembly of larger LexAOp response elements
165818	p5xQUAS	Synthetic 5xQUAS QF driver response element in enhancer grammar, requires minimal promoter for assembly
165819	pGal4BD 1-147	1-147 AA of GAL4 DNA binding domain, requires assembly with TL2-TL4 linker and TL5 activation domain
165820	pGAL4AD 768-881	768-881 AA of GAL4 activation domain, requires assembly with TL2-TL4 linker and TL1 DNA binding domain
165821	pLexA BD	LexA DNA binding domain, requires assembly with TL2-TL4 linker and TL5 activation domain
165822	pVP16 AD	VP16 activation domain, requires assembly with TL2-TL4 linker and TL1 DNA binding domain
165823	pQFBD	QF DNA binding domain, requires assembly with TL2-TL4 linker and TL5 activation domain
165824	pQFAD	QF activation domain, requires assembly with TL2-TL4 linker and TL1 DNA binding domain
165825	pGAL80	GAL4 inhibitor coding sequence (CDS)
165826	pQS	QF inhibitor coding sequence (CDS)
165827	pSV40L-TN	Viral Simian virus 40 Late poly(a) terminator element Compatible with transposon 3' ITRs
165828	pmCherry	mCherry red fluorescent protein CDS
165829	psfGFP	sfGFP green fluorescent protein CDS
165830	pEGFP	EGFP green fluorescent protein CDS
165831	pEBFP2	EBFP2blue fluorescent protein CDS
165832	pmCherry TL1	mCherry red fluorescent protein for N-terminal tagging
165833	pmCherry TL5	mCherry red fluorescent protein for C-terminal tagging
165834	pEGFP TL1	sfGFP green fluorescent protein for N-terminal tagging
165835	pEGFP TL5	sfGFP green fluorescent protein for C-terminal tagging
165836	pNPTII	G418 sulfate (geneticin) resistance marker
165837	pPAC	puromycin resistance marker
165838	pBSR	blasticidin S resistance marker
165839	pHPH	hygromycin B resistance marker
165840	pBLE	Zeocin/Phleomycin resistance marker
165841	pTK	ganciclovir sensitivity marker
165842	pFCU1	5-fluorocytosine sensitivity marker
165843	pFC31 attB O1	_C31 integrase attB site in pColE1_Omega1
165844	pFC31 attB O2	_C31 integrase attB site in pColE1_Omega2
165845	pFC31	_C31 integrase CDS
165846	pWhite_CDS	White gene CDS
165856	pUPD2	Universal Part domesticator vector v2 used for integration (domestication) of new DNA elements in the GB2.0 cloning system uses blue-white screening
165857	pVD2	universal Vector Domesticator vector v2 used in the integration (domestication) of new vector backbone parts in the GB2.0 cloning system. Uses blue-white screening method
165858	P[acman]-Alpha1	Alpha1 Basic cloning vector GB2.0 compatible P[acman]-derived backbone. Allows for copy induction from low to high using AutoFOS or similar. Requires EPI300 bacterial strain (DH10B Derived) or similar. uses blue-white screening
165859	P[acman]-Alpha2	Alpha2 Basic cloning vector GB2.0 compatible P[acman]-derived backbone. Allows for copy induction from low to high using AutoFOS or similar. Requires EPI300 bacterial strain (DH10B Derived) or similar. uses blue-white screening
165860	P[acman]-Omega1	Omega1 Basic cloning vector GB2.0 compatible P[acman]-derived backbone. Allows for copy induction from low to high using AutoFOS or similar. Requires EPI300 bacterial strain (DH10B Derived) or similar. uses blue-white screening
165861	P[acman]-Omega2	Omega2 Basic cloning vector GB2.0 compatible P[acman]-derived backbone. Allows for copy induction from low to high using AutoFOS or similar. Requires EPI300 bacterial strain (DH10B Derived) or similar. uses blue-white screening
165862	pR6Kg-Alpha1	Alpha level R6K_ origin GB Cloning vector, Conditionally replicative ori, requires pir gene expression requires EC100D or EC100D116 bacterial strain (DH10B derived) uses blue-white screening
165863	pR6Kg-Alpha2	Alpha level R6K_ origin GB Cloning vector, Conditionally replicative ori, requires pir gene expression requires pir gene expression requires EC100D or EC100D116 bacterial strain (DH10B derived) uses blue-white screening
165864	pR6Kg-Omega1	Omega level R6K_ origin GB Cloning vector, Conditionally replicative ori, requires pir gene expression requires pir gene expression requires EC100D or EC100D116 bacterial strain (DH10B derived)
165865	pR6Kg-Omega2	Omega level R6K_ origin GB Cloning vector, Conditionally replicative ori, requires pir gene expression requires pir gene expression requires EC100D or EC100D116 bacterial strain (DH10B derived) uses blue-white screening
165866	pR6Kg-Alpha1spm	Modified Alpha level R6K_ origin GB Cloning vector, Conditionally replicative ori, requires pir gene expression requires pir gene expression requires EC100D or EC100D116 bacterial strain (DH10B derived) uses blue-white screening
165867	pColE1-AlphaOmega	Terminal pBlueScript derived high-copy GB Cloning vector with both Alpha and Omega Entry points requires pir gene expression requires EC100D or EC100D116 bacterial strain (DH10B derived)uses blue-white screening
165868	pEGFP-NT-Tagging	Template for amplification of EGFP N-terminal tagging cassette
165869	pmCherry-CT-Tagging	Template for amplification of mcherry C-terminal tagging cassette
165870	pmCherry-NT-Tagging	Template for amplification of mCherry N-terminal tagging cassette
165871	pEGFP-CT-Tagging	Template for amplification of EGFP C-terminal tagging cassette
165872	pMiniwhiteA1	Basic transgenesis Mini-white physical eye marker cassette in Alpha1, lacks attB site
165873	pMiniWhiteA2	Basic transgenesis Mini-white physical eye marker cassette in Alpha2, lacks attB site
165874	pGMR-WhiteA2	Basic transgenesis 5xGMR white CDS physical eye marker cassette in Alpha2, lacks attB site
165875	p3XP3-WhiteA2	Basic transgenesis 3xP3 white CDS physical eye marker cassette in Alpha2, lacks attB site
165876	pHSP70B-CP6-nptII	Basic G418 resistance cassette in Alpha1, lacks physical marker or attB site
165877	pHSP70B-CP6-pac	Basic Puromycin resistance cassette in Alpha1, lacks physical marker or attB site
165878	pHSP70B-CP6-bsr	Basic Blasticidin resistance cassette in Alpha1, lacks physical marker or attB site
165879	pHSP70B-CP6-hph	Basic Hygromycin resistance cassette in Alpha1, lacks physical marker or attB site
165880	pHSP70B-CP6-ble	Basic Phleomycin resistance cassette in Alpha1, lacks physical marker or attB site
165881	pHSP70B-CP6-TK	Basic ganciclovir or acyclovir sensitvity cassette in Alpha2, lacks physical marker or attB site
165882	pHSP70B-CP6-FCU1	Basic 5-fluorocytosine sensitivity cassette in Alpha2, lacks physical marker or attB site
165883	pG418R-GMRWhite	G418 resistance vector with attB and eye marker. Contains an Alpha GB cloning entry point for addition of custom assembly uses blue-white screening
165884	pPuroR-GMRWhite	Puromycin resistance vector with attB and eye marker. Contains an Alpha GB cloning entry point for addition of custom assembly uses blue-white screening
165885	pBlastR-GMRWhite	Blasticidin resistance vector with attB and eye marker. Contains an Alpha GB cloning entry point for addition of custom assembly uses blue-white screening
165886	pHygroR-GMRWhite	Hygromycin resistance vector with attB and eye marker. Contains an Alpha GB cloning entry point for addition of custom assembly uses blue-white screening
165887	pRMCE{G418R-5FCS-GMRWhite}-Vasa-FC31	G418 resistant FC31 integrase mediated RMCE cassette for upgrading MiMic insertions
165888	pRMCE{PuroR-5FCS-GMRWhite}-Vasa-FC31	Puromycin resistant FC31 integrase mediated RMCE cassette for upgrading MiMic insertions
165889	pRMCE{BlastR-5FCS-GMRWhite}-Vasa-FC31	Blasticidin resistant FC31 integrase mediated RMCE cassette for upgrading MiMic insertions
165890	pPuroR-MCS-GAL4-MiniWhite	Puromycin resistant GAL4 driver vector with Mini-w+ CDS eye marker. Enhancer grammar GB20 entry point for custom enhancers. Cut-and-paste cloning can be used. Purple-white bacteria colony screening.
165891	p5xUAS	Synthetic 5xUAS GAL4 driver response element in enhancer grammar, requires minimal promoter for assembly
165894	pG418R-MCS-LexA-VP16-MiniWhite	G418 resistant LexA driver vector with Mini-w+ CDS eye marker. Enhancer grammar GB20 entry point for custom enhancers. Cut-and-paste cloning can be used. Purple-white bacteria colony screening.
165898	pPuroR-MCS-LexA-VP16-MiniWhite	Puromycin resistant LexA driver vector with Mini-white CDS eye marker. Contains an enhancer grammar GB20 entry point for custom enhancers. Standard cut-and-paste cloning can also be used. Vector uses Purple-White bacteria colony screening for identification of correct cloning
165900	pBlastR-MCS-LexA-VP16-MiniWhite	Blasticidin resistant LexA driver vector with Mini-white CDS eye marker. Contains an enhancer grammar GB20 entry point for custom enhancers. Standard cut-and-paste cloning can also be used. Vector uses Purple-White bacteria colony screening for identification of correct cloning
165902	pG418R-MCS-QF-MiniWhite	G418 resistant QF driver vector with Mini-white CDS eye marker. Contains an enhancer grammar GB20 entry point for custom enhancers. Standard cut-and-paste cloning can also be used. Vector uses Purple-White bacteria colony screening for identification of correct cloning
165906	pBlastR-MCS-QF-MiniWhite	Blasticidin resistant QF driver vector with Mini-white CDS eye marker. Contains an enhancer grammar GB20 entry point for custom enhancers. Standard cut-and-paste cloning can also be used. Vector uses Purple-White bacteria colony screening for identification of correct cloning
165907	pBlastR-5xUAS-sfGFP-GMRWhite	Blasticidin resistant 5xUAS sfGFP GMR-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165908	pBlastR-5xUAS-mCherry-GMRWhite	Blasticidin resistant 5xUAS mCherry GMR-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165910	pBlastR-5xUAS-sfGFP-MiniWhite	Blasticidin resistant 5xUAS sfGFP Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165911	pBlastR-5xUAS-mCherry-MiniWhite	Blasticidin resistant 5xUAS mCherry Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165912	pBlastR-5xUAS-EBFP2-MiniWhite	Blasticidin resistant 5xUAS EBFP2 Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165916	pBlastR-12xLexAOp-sfGFP-MiniWhite	Blasticidin resistant 12xLexAOp sfGFP Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165917	pBlastR-12xLexAOp-mCherry-MiniWhite	Blasticidin resistant 12xLexAOp mCherry Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165918	pBlastR-12xLexAOp-EBFP2-MiniWhite	Blasticidin resistant 12xLexAOp EBFP2 Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165920	pBlastR-5xQUAS-mCherry-GMRWhite	Blasticidin resistant 5xQUAS mCherry GMR-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165921	pBlastR-5xQUAS-EBFP2-GMRWhite	Blasticidin resistant 5xQUAS EBFP2 GMR-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165922	pBlastR-5xQUAS-sfGFP-MiniWhite	Blasticidin resistant 5xQUAS sfGFP Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165923	pBlastR-5xQUAS-mCherry-MiniWhite	Blasticidin resistant 5xQUAS mCherry Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening
165924	pBlastR-5xQUAS-EBFP2-MiniWhite	Blasticidin resistant 5xQUAS EBFP2 Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening