The Nucleus v0.1.0 distribution plate is the first release of the physical DNA encoding Nucleus content components and modules, available under the OpenMTA.

Table of Contents

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Backbone	pOpen v3
Antibiotic Resistance	Amp
Concentration (fmol)	10
Total volume (ul)	10
Sequence Information	[Github]

Plate Map

	1	2	3	4	5	6	7	8	9	10	11	12	Category
A	AlaRS	ArgRS	AsnRS	AspRS	CysRS	GlnRS	GluRS	GlyQS-DualHis	HisRS	IleRS	LeuRS	LysRS	PURE
B	MetRS	PheST-DualHis	ProRS	SerRS	ThrRS	TrpRS	TyrRS	ValRS	MTF	IF1	IF2	IF3	PURE
C	EF-G	EF-Tu	EF-Ts	RF1	RF2	RF3	RRF	AK-Gg	CK-Gg	NDK	PPiase	T7RNAP	PURE
D	GlyS	GlyQ	GlySQ	GlyQS	PheS	PheT	PheST	PheTS	AK-Sc-1	AK-Sc-2	AK-Oc	AK-Ec	PURE
E	CK-Oc	plamGFP-Chimeric	plamGFP-PURE	plamGFP-TXTL	mmilCFP	meleRFP	cjBlue	cjBlue-lacO	cjBlue-lacO-His6	eforRed	eforRed-lacO	eforRed-lacO-His6	PURE / Rep
F	PURET7-1	PURET7-2	PURET7-3	PURET7-4	PURET7-5	PURET7-6	PURET7-7	PURET7-8	PURET7-9	PURET7-10	amajLime	gfasPurple	T7 / Rep
G	tetR	lacI	pT7-tetO	pT7-lacO	pT7-tetO-lacO	pT7-lacO-tetO							Module
H	UTR1	RBS	tT7	tT7hyb6	tT7hyb10	pOpenv3-MCL0							MoClo

The content of the plate is described in the DNA Distribution document. In general, the plate is arrayed by plasmid category:

PURE genes occupy the first three rows (A-C), in the gene ordering originally published by Shimizu, et al (2001).
Row D contains alternative versions of the PURE genes, such as different designs for the two-subunit tRNA synthetases, and metabolic enzymes sourced from different organisms as have been used in the literature.
Row E contains open measurement reporters.
Row F contains T7 promoter variants engineered to span two orders of magnitude of expression, for direct testing or use as a MoClo level 0 promoter part.
Row G contains inducible modules for use in the PURE cell free system.
Row H contains several basic MoClo level 0 parts for assembly of level 1 transcription units.

Technical Information

All DNA is encoded on the high-copy pOpen v3 plasmid, in the Nucleus DNA architecture.
All plasmids on the plate are Ampicillin / Carbenicillin resistant.
Each construct is normalized to 10 fmol / uL and provided as 10 uL total.
Plates are sealed with slit silicon plate seals and lids. DNA can be extracted through the slits in the seal without removing it.
- Multichannel pipetting through the slit seal is very difficult.
- Exercise caution if removing the seal—be very gentle, or the vacuum produced will pull sample from the well and potentially cross-contaminate the wells.
DNA may have shifted in transit—spin down the plate before using it in downstream work.

Getting Started

We recommend sub-cloning the DNA into an appropriate cloning strain such as DH5-alpha or NEB Stable, and preparing more high concentration DNA for downstream use.

Spin down the plate in a tabletop plate spinner, or a centrifuge at 2000g for 1 minute.