Plasmids

The entire collection of plasmids, distributed as live cultures on two plates in a 96-well format, will be priced at $400 for academic/non-profit users and $800 for commercial/industry. Individual plasmids from the kit can also be ordered as bacterial cultures on agar plugs for standard plasmid pricing. You may view the individual list of plasmids in this collection here: https://www.chlamycollection.org/products/moclo-toolkit/. 

From Crozet et al. (2018): “… we have developed a Modular Cloning toolkit for the green microalga Chlamydomonas reinhardtii. It is based on Golden Gate cloning with standard syntax, and comprises 119 openly distributed genetic parts, most of which have been functionally validated in several strains. It contains promoters, UTRs, terminators, tags, reporters, antibiotic resistance genes, and introns cloned in various positions to allow maximum modularity. The toolkit enables rapid building of engineered cells for both fundamental research and algal biotechnology. This work will make Chlamydomonas the next chassis for sustainable synthetic biology.”

MoClo Kit Sequences

MoClo Kit Documents

Crozet et al. (2018) Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synthetic Biology 7:2074–2086

We are sorry but we cannot ship the MoClo Toolkit to China because of long shipping and customs delays. Individual clones can still be shipped however. 

From Jean-David Rochaix to Boynton-Gillham laboratory, 1981

Chloroplast DNA BamH1 14/15 (3.0 kb) from Rochaix’s cw15 strain in pBR313; contains 3′ end of 23S rRNA gene through 5S; see P-124 for same insert in pBR322

host strain: C600
amp resistant

More information

Strain: Rochaix cw15

Insert: chloroplast DNA BamHI 14/15 (3.0 kb)

Genes present: 23S rRNA (3′ end), 5S rRNA

Vector: pBR313

Bacterial host strain: C600

Selectable marker: amp-r tet-s

Origin: Single cell clone of P-3 (Ba1 from Rochaix received 6/81)

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

From Jean-David Rochaix to Boynton-Gillham laboratory, 1981

Chloroplast DNA BamHI 10 (7.6 kb) from Rochaix’s cw15 strain in pBR313; contains entire atpB gene and 3′ portion of ORF1995

host strain: C600
amp resistant

More information

Strain: Rochaix cw15

Insert: chloroplast DNA BamHI 10 (7.6 kb)

Genes present: atpB

Vector: pBR313

Bacterial host strain: C600

Selectable marker: amp-r tet-s

Origin: Single cell clone of P-6 (Ba 5 from Rochaix, received 6/81)

Reference: Boynton et al. (1988). See for use of this plasmid in transformation of atpB gene, and analysis of 12 bp tandem repeats./p>

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

From Jean-David Rochaix to Boynton-Gillham laboratory, 1981

Chloroplast DNA BamHI 8 (9.6 kb) from Rochaix’s cw15 strain in pBR313; contains most of rpoC2 (3′) and 5′ end of psaB; see P-122 for same insert in pBR322

host strain: C600
amp resistant

Additional Information

Strain: Rochaix cw15

Insert: chloroplast DNA BamHI 8 (9.6 kb)

Genes present: rpoC2 (3′ end), psaB (extreme 5′ end), ARS04,02

Vector: pBR313

Bacterial host strain: C600

Selectable marker: amp-r tet-s

Origin: Single cell clone of P-8 (Ba7 from Rochaix, received 6/81)

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

From Jean-David Rochaix to Boynton-Gillham laboratory, 1981

Chloroplast DNA BamHI 6 (13.4 kb) from Rochaix’s cw15 strain in pBR313; contains 3′ end of rps2 rpoA, psbB, psbN, and psbH; P-202 is the same insert in pBR322

host strain: C600
amp resistant

More information

Strain: Rochaix cw15

Insert: chloroplast DNA BamHI 6 (13.4 kb)

Genes present: psbH, psbN, ORF31, psbB, probably rps2

Vector: pBR313

Bacterial host strain: C600

Selectable marker: amp-r tet-s

Origin: Single cell clone of P-10 (Ba10 from Jean-David Rochaix, received 6/81

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA BamHI 9 (8.4 kb) in pUC8, from CC-125 wild type; contains rpl14, rps8, psaA exon 1, and rps4

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA BamHI 9 (8.4 kb)

Genes present: rpl14, rpl5, rps8, psaA (exon 1), rps4

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA BamHI 11/12 (7.0 kb) in pUC8, from CC-125 wild type; contains 16S rRNA through 23S 5′ end

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA BamHI 11/12 (7.0 kb)

Genes present: 16S rRNA, 23s rRNA (5′ end)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

See P-271 for the same insert in pBR322.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 5 (10.5 kb) in pUC8, from CC-125 wild type; contains psbB, rpoA, rps2 and most of rps18

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 5 (10.5 kb)

Genes present: psbB (5′), rps2, atpF

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Jeff Palmer in Boynton/Gillham lab, 1986

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 8 (8.5 kb) in pUC8, from CC-125 wild type; contains rps8, psaA exon 1, rps4 and 5′ end of 16S rRNA

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 8 (8.5 kb)

Genes present: rps8, psaA (exon 1), rps4, 16S rRNA (5′ end)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 9 (7.6 kb) in pUC8, from CC-125 wild type; contains clpP, rpl35, rpl23, and 5′ half of rpl2

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 9 (7.6 kb)

Genes present: clpP, trnL, petB, chlL, rpl23, rpl2 (5′)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 12 (6.5 kb) in pUC8, from CC-125 wild type; contains psaA exon 3

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 12 (6.5 kb)
Genes present: psaA (exon 3)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 15 (6.0 kb) in pUC8, from CC-125 wild type; contains psbA exons 1-4

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 15 (5.8 kb)psbA

Genes present: psbA exons 1-3

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 14 (5.8 kb) in pUC8, from CC-125 wild type; contains psaB, rbcL, and 5′ end of atpA

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 14 (5.5 kb)

Genes present: psaB, rbcL, atpA (5′)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 15 (5.8 kb) in pUC8, from CC-125 wild type; contains psbA exons 1-4

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 15 (5.8 kb) Gene psbA

Genes present: psbA (exons 1, 2, 3)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

Comment Same insert as P-66, but opposite orientation.

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 16 (5.0 kb) in pUC8, from CC-125 wild type; contains 3′ end of rpl2, rps19, rpl16, rpl14, and rpl5

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 16 (5.0 kb)

Genes present: rpl2 (part), rps19, rpl16, oriA, rpl14, rpl5

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 17 (4.9 kb) in pUC8, from CC-125 wild type; contains rps12, psaJ, atpI, and psbJ

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 17 (4.9 kb)

Genes present: psaJ, rps12

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 18 (4.9 kb) in pUC8, from CC-125 wild type; contains rps11, tscA, and most of chlN

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert:

Genes present:

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 19 (4.4 kb) in pUC8; contains psbH, psbN, and psbT

host strain: JM83
amp resistant

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 24 (3.3 kb) in pUC8, from CC-125 wild type; contains ycf12 and atpE

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 24 (3.3 kb)

Genes present: atpE

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab

Comment The 350 bp EcoRI-Bgl II subclone of this plasmid is a good probe for atpE.

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 26 (3.0 kb) in pUC8, from CC-125 wild type; internal to ORF2971

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 26 (3.0 kb)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 31 (1.4 kb) in pUC8, from CC-125 wild type; internal to rpoC2

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 31 (1.4 kb)

Genes present: rpoC2 (part)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 32 (1.3 kb) in pUC8, from CC-125 wild type; contains 3′ half of tufA

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 32 (1.3 kb)

Genes present: tufA (3′ end)

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA PstI 4 (15.1 kb) in pBR322, from CC-125 wild type; contains rps2, rps18, ycf3, ycf4, rps9, psbE, rpoB and psbF

host strain: SK2267
tetracycline resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA PstI 4 (15.1 kb)

Genes present: rps2 (probably), atpF, psbE, rpoB, psbF

Vector: pBR322

Bacterial host strain: SK2267

Selectable marker: tet-r, amp-s

Origin: David Lamb in Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Orientation of P-78

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA PstI 14 (3.8 kb) in pBR322, from CC-125 wild type; contains psbT and psbB

host strain: SK2267
tetracycline resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA PstI 14 (3.8 kb)

Genes present: psbB

Vector: pBR322

Bacterial host strain: SK2267

Selectable marker: tet-r, amp-s

Origin: David Lamb in Boynton/Gillham lab, 1983

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA PstI 19 (2.9 kb) in pBR322, from CC-125 wild type; contains rpoC1b

host strain: SK2267
tetracycline resistant

Boynton-Gillham laboratory, Duke University, 1983

Mitochondrial Bam-Sal fragment (5.6 kb) in pBR322, from CC-125 wild type; contains cox1, nd2, nd6, W, L6, L7, Q, and S1

host strain: SK2267
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: mitochondrial DNA BamHI-SalI (5.6 kb)

Genes present: cox1, nd2, nd6, W, L6, L5, L7, Q, S1

Vector: pBR322

Bacterial host strain: SK2267

Selectable marker: amp-r

Origin: Boynton/Gillham lab, 1983

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA EcoRI 22 (3.5 kb) in pUC8, from CC-125 wild type; contains nearly all of atpA, psbI, and most of cemA

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA EcoRI 22 (3.5 kb)

Genes present: atpA (most of gene; Eco site is near the 5′ end); psbI

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Boynton-Gillham lab, 1983

Comment: This is a good probe for atpA.

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Chloroplast DNA BamHI 7 (11.6 kb) in pUC8, from CC-125 wild type; contains part of the Wendy transposon, petA, petD, and chlB

host strain: JM83
amp resistant

More information

Strain: CC-125 wild type mt+ 137c

Insert: chloroplast DNA BamHI 7 (11.6 kb)

Genes present: petA, petD, chlB

Vector: pUC8

Bacterial host strain: JM83

Selectable marker: amp-r

Origin: Boynton/Gillham lab, 1985

This map is from the original file for the plasmid and was drawn before the complete sequence was known.

Boynton-Gillham laboratory, Duke University, 1983

Nuclear rDNA unit, 3.6 kb BamH1 fragment in pUC8, from CC-125 wild type; contains part of 25S, 5.8S, and most of 18S rDNA

For map, see Marco and Rochaix (1980) Mol. Gen. Genet. 177, 715-723

host strain: JM83
amp resistant

Marco Y, Rochaix JD (1980) Organization of the nuclear ribosomal DNA of Chlamydomonas reinhardii. Mol Gen Genet 177:715-23

From Jean-David Rochaix to Boynton-Gillham laboratory, 1985

Chloroplast DNA EcoRI 27 (2.7 kb) from Rochaix’s cw15 strain in pCR1; contains most of psbD and the 5′ end of ORF2971

host strain: JM83
kanamycin resistant

More information

Strain: Rochaix cw15

Insert: chloroplast DNA EcoRI 27 (2.7 kb)

Genes present: psbD

Vector: pCR1 (11.4 kb)

Bacterial host strain: Uncertain

Selectable marker: kan-r

Origin: Jean-David Rochaix, 1985, as R3

This map is from the original file for the plasmid and was drawn before the complete sequence was known.