Golden Gate Entry Clones

The following are insert clones for SapI Golden Gate clones. The plasmids are organized based on slots (SapI overhangs of the insert clone). All clones encoding proteins have the reading frame of the 3 base overlap.

Sequences for plasmids described in Knoebel et al. 2023:

Knoebel et al. 2023 plasmids [all seqs]

Sequence of individual clone by slot position:

AAC TGG slot clones

NM #	Insert	SapI sites	.gb file [all seqs]
3851	rps-13 3′	AAC TGG	[seq]
4099	lexO 5X	AAC TGG	[seq]
4335	QUAS 5X	AAC TGG	[seq]

5′ arm slot clones

NM #	Insert	SapI sites	.gb file [all seqs]
3700	11X UAS	TGG GCG	[seq]
3703	5X lexO	TGG GCG	[seq]
3704	5X QUAS	TGG GCG	[seq]
3770	7X tetO	TGG GCG	[seq]
3909	7X tetO ∆mec-7p	TGG GCG	[seq]
4035	tiTT4348 left arm	TGG GCG	[seq]
4053	cxTi10882 left arm	TGG GCG	[seq]
4145	7X tetO ∆pes-10p	TGG GCG	[seq]
4260	oxTi365 left arm	TGG GCG	[seq]

5′ arm-CT promoter clones

NM #	Insert	SapI sites	.gb file [all seqs]
3695	tetO 7x  ∆pes-10p	TGG ATG	[seq]
3837	rab-3p long	TGG ATG	[seq]
3858	glr-1p	TGG ATG	[seq]
3863	mec-4 syn p	TGG ATG	[seq]
3896	ehs-1p	TGG ATG	[seq]
3899	tetO 7X  ∆mec-7p	TGG ATG	[seq]
3932	dpy-7p	TGG ATG	[seq]
3936	F49H12.4p	TGG ATG	[seq]
3938	cup-4p	TGG ATG	[seq]
3945	rpl-27p	TGG ATG	[seq]
3947	nhx-2p	TGG ATG	[seq]
3985	myo-2p	TGG ATG	[seq]
4037	myo-3p	TGG ATG	[seq]
4104	myo-3p	TGG ATG	[seq]
4106	pgp-12p	TGG ATG	[seq]
4117	pgp-12p	TGG ATG	[seq]
4119	unc-25p	TGG ATG	[seq]
4303	lys-8p	TGG ATG	[seq]

CT slot clones

NM #	Insert	SapI sites	.gb file [all seqs]
3401	linker	GCG ATG	[seq]
3495	3X HA	GCG ATG	[seq]
3496	3X FLAG	GCG ATG	[seq]
3524	GSGA P2A	GCG ATG	[seq]
3706	∆pes-10p	GCG ATG	[seq]
3767	∆mec-7p	GCG ATG	[seq]
4096	3X lexO ∆pes-10p	GCG ATG	[seq]
4232	11X UAS ∆pes-10p	GCG ATG	[seq]
4341	5X QUAS ∆pes-10p	GCG ATG	[seq]

CT-NT Clones

NM #	Insert	SapI sites	.gb file [all seqs]
4466	myo-2p FRT cyOFP FLP mNG	GCG ACG	[seq]
4630	7X GFP11 loxP hygR loxP	GCG ACG	[seq]
4697	mNG hygR	GCG ACG	[seq]

FP slot clones

NM #	Insert	SapI sites	.gb file [all seqs]
3460	TIR1	ATG AAG	[seq]
3578	3X HA turboID	ATG AAG	[seq]
3686	tdTomato	ATG AAG	[seq]
3699	mNeonGreen	ATG AAG	[seq]
3821	tetR-L-QF (tetOFF)	ATG AAG	[seq]
3943	ctns-1	ATG AAG	[seq]
4045	tetR-LL-QF	ATG AAG	[seq]
4064	nls GFP-C1 nls	ATG AAG	[seq]
4067	cyOFP	ATG AAG	[seq]
4068	nls cyOFP	ATG AAG	[seq]
4137	mScarlet	ATG AAG	[seq]
4226	tagBFP	ATG AAG	[seq]
4233	GFP1-10-C1	ATG AAG	[seq]
4241	FLP D5	ATG AAG	[seq]
4259	tomm-20	ATG AAG	[seq]
4338	QF2	ATG AAG	[seq]
4339	lexA-L-QF	ATG AAG	[seq]
4340	Gal4-L-QF	ATG AAG	[seq]
4464	nls CRE	ATG AAG	[seq]
4491	nls scarlet	ATG AAG	[seq]
4492	nls mNeonGreen	ATG AAG	[seq]
4493	nls BFP	ATG AAG	[seq]
4503	7X GFP11	ATG AAG	[seq]

SEC slot clones

NM #	Insert	SapI sites	.gb file [all seqs]
3432	loxP	AAG GGT	[seq]
3928	gpd-2/3 3′ UTR	AAG GGT	[seq]
4114	V2A	AAG GGT	[seq]
4299	3X HA	AAG GGT	[seq]
4300	3X FLAG	AAG GGT	[seq]
4301	flex linker	AAG GGT	[seq]
4302	rigid linker	AAG GGT	[seq]
4602	loxP hygR	AAG GGT	[seq]

SEC-3′ arm slot clones

NM #	Insert	SapI sites	.gb file [all seqs]
3694	tbb-2 3′ UTR	AAG GTA	[seq]
3751	act-4 3′ UTR	AAG GTA	[seq]
3760	unc-54 3′ UTR	AAG GTA	[seq]
3766	let-858 3′ UTR	AAG GTA	[seq]
4215	glh-2 3′ UTR	AAG GTA	[seq]

NT slot clones

NM #	Insert	SapI sites	.gb file [all seqs]
3402	flex linker	GGT ACG	[seq]
3407	T2A peptide	GGT ACG	[seq]
3413	AID	GGT ACG	[seq]
3503	3X FLAG	GGT ACG	[seq]
3527	rab-3	GGT ACG	[seq]
3887	GFP-C1	GGT ACG	[seq]
3911	scarlet-I-C1	GGT ACG	[seq]
3980	mNeonGreen	GGT ACG	[seq]
4127	nls cyOFP nls	GGT ACG	[seq]
4138	nls mScarlet-C1 nls	GGT ACG	[seq]
4139	nls mNeonGreen nls	GGT ACG	[seq]
4230	nls tag-BFP-C1	GGT ACG	[seq]
4427	nls-GFP-C1	GGT ACG	[seq]
4502	7X GFP11	GGT ACG	[seq]
4847	myr-tagRFP-T	GGT ACG	[seq]

3′ arm slot clones

NM #	Insert	SapI sites	.gb files [all seqs]
3777	tbb-2 3′ UTR	ACG GTA	[seq]
3822	act-4 3′ UTR	ACG GTA	[seq]
3979	let-858 3′ UTR	ACG GTA	[seq]
3981	glh-2 3′ UTR	ACG GTA	[seq]
4036	ttTi4348 right arm	ACG GTA	[seq]
4054	cxTi10882 right arm	ACG GTA	[seq]
4261	oxTi365 right arm	ACG GTA	[seq]
4313	rps-17 3′ UTR	ACG GTA	[seq]

Clones organized by use:

HygR Cassettes

NM #	Insert	SapI sites	.gb file
4602	loxP hygR loxP	AAG GGT	[seq]
4630	7X GFP11 loxP hygR loxP	GCG ACG	[seq]
4697	mNG loxP hygR loxP	GCG ACG	[seq]

landing site tools

NM #	Insert	SapI sites	.gb files
4035	tiTT4348 left arm	TGG GCG	[seq]
4053	cxTi10882 left arm	TGG GCG	[seq]
4260	oxTi365 left arm	TGG GCG	[seq]
4036	ttTi4348 right arm	ACG GTA	[seq]
4054	cxTi10882 right arm	ACG GTA	[seq]
4261	oxTi365 right arm	ACG GTA	[seq]
4466	myo-2p FRT cyOFP FLP mNG	GCG ACG	[seq]

References:

Knoebel, E., Dour, S., & Nonet, M. L. (2023). A toolkit for assembly of targeting clones for C. elegans transgenesis. microPublication Biology. 10.17912/micropub.biology.000966.

Last updated: 10-2-2023