OVC Core and Extended Collections
(Kit # 1000000164, 1000000165 )

Depositing Lab:   Harald Janovjak

The Janovjak Lab has provided both a Core and Extended Optogenetic Vector Collection (OVC).

Kit #1000000164

The Core Collection contains in total 24 plasmids: three plasmids containing the empty cloning cassette in configurations ABC, BAC or ACB, and 21 plasmids containing one of nine light-sensing domains (LSDs) or LSD binding partners in A as well as separately in site C (N- and C-terminally, respectively).

Kit #1000000165

The Extended Collection contains in total 17 plasmids: Four plasmids containing the cloning cassette with alternative linker configurations in site B, eight plasmids containing one of two LSDs engineered to heterodimerize and their binding partners, and five plasmids containing different constructs of a light-activated Opto-caspase 9.

These kits will be sent as bacterial glycerol stocks in 96-well plate format.

Original Publication

Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein-Protein Interactions Tichy A-M, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak H. J Mol Biol.2019 Aug 9;431(17):3046-3055. doi: 10.1016/j.jmb.2019.05.033. PubMed PMID: 31150735.

Description

The Optogenetic Vector Collection (OVC) contains two sets of plasmids (the Core and the Extended Collection) to rapidly generate custom-made optogenetic tools for the manipulation of protein-protein interactions. The vectors are based on a modular cloning cassette (the primary cassette is termed ABC) containing three insertion sites (A, B, and C). Restriction sites A and C (AgeI and XmaI, respectively) generate compatible cohesive ends so that the target sequence can be inserted both N- or C-terminally using only one PCR product.

OVC Core Collection: (A) ABC and alternative cassettes. Cassettes include Kozak sequence, start codon and stop codon. (B) Cloning strategy based on the vector collection allows universal target amplicon insertion both N- and C-terminally. (C) LSDs and binding partners included in the OVC Core Collection.

OVC Extended Collection: (A) ABC cassettes with alternative linkers. (B) Heterodimerizing LSDs based on AsLOV2 and their binding partners (C) Opto-casp9 constructs with N- and C-terminal fusion of the LSD to the casp9 domain included in the OVC Extended Collection.

Kit Documentation

Workflow 192.9 KB

How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:

“OVC Core Collection was a gift from Harald Janovjak (Addgene kit #1000000164).”

“OVC Extended Collection was a gift from Harald Janovjak (Addgene kit #1000000165).”

For your Reference section:

Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein-Protein Interactions Tichy A-M, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak H. J Mol Biol.2019 Aug 9;431(17):3046-3055. doi: 10.1016/j.jmb.2019.05.033. PubMed PMID: 31150735.

OVC Core Collection - #1000000164

Inventory

Well	Plasmid	Resistance
A / 1	pOVC1	Ampicillin
A / 2	pOVC2	Ampicillin
A / 3	pOVC3	Ampicillin
A / 4	pOVC1_VfAU1-LOV_X	Ampicillin
A / 5	pOVC1_CrPH-LOV_X	Ampicillin
A / 6	pOVC1_NcVVD-LOV_X	Ampicillin
A / 7	pOVC1_RsLP-LOV_X	Ampicillin
A / 8	pOVC1_MxCarH-CBD_X	Ampicillin
A / 9	pOVC1_TtCarH-CBD_X	Ampicillin
A / 10	pOVC1_AtCRY2-PHR_X	Ampicillin
A / 11	pOVC1_AtCIB_X	Ampicillin
A / 12	pOVC1_AtPIF6_X	Ampicillin
B / 1	pOVC1_AtPHYB-S_X	Ampicillin
B / 2	pOVC1_ScPH1-S_X	Ampicillin
B / 3	pOVC1_X_VfAU1-LOV	Ampicillin
B / 4	pOVC1_X_CrPH-LOV	Ampicillin
B / 5	pOVC1_X_NcVVD-LOV	Ampicillin
B / 6	pOVC1_X_RsLP-LOV	Ampicillin
B / 7	pOVC1_X_MxCarH-CBD	Ampicillin
B / 8	pOVC1_X_TtCarH-CBD	Ampicillin
B / 9	pOVC1_X_AtCRY2-PHR	Ampicillin
B / 10	pOVC1_X_AtPIF6	Ampicillin
B / 11	pOVC1_X_ScPH1-S	Ampicillin
B / 12	pOVC1_X_AtCIB	Ampicillin

Data calculated @ 2024-11-21

OVC Extended Collection - #1000000165

Resistance Color Key

Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.

Inventory

Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.

Kit Plate Map

96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.

Resistance Color Key

Ampicillin

Inventory

Well	Plasmid	Resistance
A / 1	pOVC1_L1(GSSSG)	Ampicillin
A / 2	pOVC1_L2(GSSGGSSG)	Ampicillin
A / 3	pOVC1_L3(GSSSSGGSSSG)	Ampicillin
A / 4	pOVC1_L4(EAAAK)3	Ampicillin
A / 5	pOVC1_EcSSPB-nano_X	Ampicillin
A / 6	pOVC1_EcSSPB-micro_X	Ampicillin
A / 7	pOVC1_HsPDZ1b_X	Ampicillin
A / 8	pOVC1_X_AsPT1-LOV2-pep	Ampicillin
A / 9	pOVC1_X_HsPDZ1b	Ampicillin
A / 10	pOVC1_X_AsPT1-LOV2-EcSsra	Ampicillin
A / 11	pOVC1_X_EcSSPB-micro	Ampicillin
A / 12	pOVC1_X_EcSSPB-nano	Ampicillin
B / 1	Opto-casp9_VfAU1-LOV_casp9	Ampicillin
B / 2	Opto-casp9_AtCRY2-PHR-casp9	Ampicillin
B / 3	Opto-casp9_casp9-VfAU1-LOV	Ampicillin
B / 4	Opto-casp9_casp9-AtCRY2-PHR	Ampicillin
B / 5	VfAU1-LOV_P2A_casp9	Ampicillin

Data calculated @ 2024-11-21