Chromoprotein kits
(Kit # 1000000174, 1000000198 )

Depositing Lab:   Anthony Forster

Kit #1000000174 is a synthetic biology collection of plasmids for expression of chromoproteins in bacteria, which can be used as an educational resource.

Kit # 1000000198 is updated from kit 1.0 to contain two new chromoprotein genes that are less toxic to E. coli so that the plasmids are more stable during growth (Bao et al., 2020).

These kits will be sent as individual bacterial stabs at room temperature.

Original Publication

Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology Liljeruhm J, Funk SK, Tietscher S, Edlund AD, Jamal S, Wistrand-Yuen P, Dyrhage K, Gynnå A, Ivermark K, Lövgren J, Törnblom V, Virtanen A, Lundin ER, Wistrand-Yuen E, Forster AC. J Biol Eng. 2018 May 10;12:8. doi: 10.1186/s13036-018-0100-0. PubMed Article .

Additional Publication for the Chromoprotein Kit 2.0

Overcoming chromoprotein limitations by engineering a red fluorescent protein. Bao L, Menon PNK, Liljeruhm J, Forster AC. Anal Biochem. 2020 Dec 15;611:113936. doi: 10.1016/j.ab.2020.113936. PubMed Article .

Description

Kit #1000000174 Chromoprotein kit 1.0

This collection deposited by Anthony Forster's lab contains a set of 11 Biobrick™ genetic modules for standardized 3A assembly. It comprises plasmids for a published teaching course (see the Protocols & Resources tab) where successful coupling of two modules causes visible chromoprotein expression in E. coli. Also included are plasmids that constitutively express other chromoproteins.

Kit #1000000198 Chromoprotein kit 2.0

This collection deposited by Anthony Forster's lab contains a set of 9 Biobrick™ genetic modules for standardized 3A assembly. It comprises plasmids for a teaching course (Liljeruhm et al., 2014), where successful coupling of two modules causes visible chromoprotein expression in E. coli. Also included are plasmids that constitutively express other useful chromoprotein genes (Liljeruhm et al., 2018).

Biobrick™ “3A” assembly standard. Image based on Fig. 25 of Liljeruhm J, Gullberg E & Forster AC (2014). Synthetic biology: a lab manual. Lab Manual

Additional Protocols

Synthetic biology: a lab manual. Liljeruhm J, Gullberg E, Forster AC. World Scientific Press (Singapore) 2014, 204 pp. doi: 10.1142/9061. Lab Manual

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:

“Chromoprotein kit 1.0 or Chromoprotein kit 2.0 was made available via the Forster lab (Addgene kit #1000000174 or #1000000198).”

For your Reference section:

Kit #1000000174: Chromoprotein kit 1.0

Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology Liljeruhm J, Funk SK, Tietscher S, Edlund AD, Jamal S, Wistrand-Yuen P, Dyrhage K, Gynnå A, Ivermark K, Lövgren J, Törnblom V, Virtanen A, Lundin ER, Wistrand-Yuen E, Forster AC. J Biol Eng. 2018 May 10;12:8. doi: 10.1186/s13036-018-0100-0. PubMed Article .

Kit #1000000198: Chromoprotein kit 2.0

Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology Liljeruhm J, Funk SK, Tietscher S, Edlund AD, Jamal S, Wistrand-Yuen P, Dyrhage K, Gynnå A, Ivermark K, Lövgren J, Törnblom V, Virtanen A, Lundin ER, Wistrand-Yuen E, Forster AC. J Biol Eng. 2018 May 10;12:8. doi: 10.1186/s13036-018-0100-0. PubMed Article .

Overcoming chromoprotein limitations by engineering a red fluorescent protein. Bao L, Menon PNK, Liljeruhm J, Forster AC. Anal Biochem. 2020 Dec 15;611:113936. doi: 10.1016/j.ab.2020.113936. PubMed Article .

The Chromoprotein kit 1.0 contains 11 plasmids, sent as bacterial stabs of the E. coli strain DH5alpha, found in the four categories below. Please refer to the individual plasmid pages below for more details on each plasmid in this kit:

Promoter (including RFP reporter)

• promoter J23110 (upstream of RBS, mRFP1)

Chromoprotein gene (including RBS, not promoter)

• asPink chromoprotein promoter-less
• amilCP chromoprotein promoter-less
• amilGFP chromoprotein promoter-less

Chromoprotein expression plasmids (including promoter and RBS in pSB1C3 vector)

• aeBlue chromoprotein
• amajLime chromoprotein
• tsPurple chromoprotein
• amilCP_Orange chromoprotein
• amilCP_Pink chromoprotein

Vector backbone (including mRFP1E reporter)

• pSB1K3 containing mRFP1
• pSB3K3 containing mRFP1

Chromoprotein kit 1.0 bacterial stabs.

The Chromoprotein kit 2.0 contains 9 plasmids, sent as bacterial stabs of the E. coli strain DH5alpha, found in the four categories below. Please refer to the individual plasmid pages below for more details on each plasmid in this kit:

Promoter (including mRFP1E reporter and Ampicillin resistance)

• promoter J23110 (upstream of RBS, mRFP1)

Chromoprotein gene (including RBS and Chlorampenicol resistance, not promoter)

• asPink chromoprotein promoter-less
• promoter-less mRFP1_Violet chromoprotein
• promoter-less mRFP1_Orange chromoprotein

Chromoprotein expression plasmids (including promoter, RBS, and Chloramphenicol resistance in pSB1C3 vector)

• amajLime chromoprotein
• aeBlue chromoprotein
• amilCP chromoprotein

Vector backbone (including mRFP1E reporter and Kanamycin resistance)

• pSB1K3 containing mRFP1
• pSB3K3 containing mRFP1

Chromoprotein kit 2.0 bacterial stabs.