CIDAR MoClo Vector Expansion Kit
(Kit #
1000000279
)
Depositing Lab: Wei Huang
This kit contains a set of low- (p15A) and medium-copy (pBR322) destination vectors ccompatible with the Cross-disciplinary Integration of Design Automation Research (CIDAR) MoClo standard. The kit also includes chloramphenicol-resistant Level 1 vectors.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
Construction and Characterization of MoClo-Compatible Vectors for Modular Protein Expression in E. coli. Nielsen JR, Lewis MJ, Huang WE. ACS Synth Biol. 2025 Jan 12. doi: 10.1021/acssynbio.4c00564. PubMed (Link opens in a new window) Article (Link opens in a new window)
Description
The CIDAR MoClo system enables standardized one-pot assembly of (multiple) genetic constructs using Golden Gate Assembly (GGA), relying on BsaI and BbsI restriction enzymes. Existing MoClo kits are often limited in their offering of destination vectors, usually requiring the user to express the construct in high-copy destination vectors. The use of low- and medium copy destination vectors can be desirable for expressing burdensome proteins, large constructs, or for deploying multi-plasmid systems.
This kit includes 28 low- and medium-copy destination vectors (p15A- and pBR322-origins of replication, respectively) to aid the user in assembling expression constructs with suitable copy numbers. Since the p15A origin of replication is compatible with both pBR322- and pUC19 (high-copy) replication origins, this kit allows the user to construct multi-plasmid systems with ease using GGA. Besides offering flexibility regarding copy number, this kit contains Level 1 destination vectors conferring chloramphenicol resistance, expanding the set of markers available for selection as found in the original CIDAR MoClo kit.
The CIDAR MoClo Vector Expansion Kit is intended for users interested in refining protein expression levels in bacterial hosts (most notably E. coli), and for those interested in using multi-plasmid systems. The kit is fully compatible with the original CIDAR MoClo Parts Kit (Kit #1000000059) deposited by Douglas Densmore, as well as MoClo-compatible parts and constructs found in later iterations of the standard.
Figure 1: The CIDAR MoClo Vector Expansion kit contains 28 destination vectors with either low- (p15A) or medium-copy (pBR322) origins of replication, allowing lower-copy plasmid expression compared to the standard pUC19-derived destination vectors. Level 1 destination vectors with either chloramphenicol or kanamycin resistance cassettes are available to offer flexibility in assembly and to facilitate multi-plasmid protein expression.
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:
"The CIDAR MoClo Vector Expansion Kit was a gift from Wei Huang (Addgene kit #1000000279)."
For your Reference section:
Construction and Characterization of MoClo-Compatible Vectors for Modular Protein Expression in E. coli. Nielsen JR, Lewis MJ, Huang WE. ACS Synth Biol. 2025 Jan 12. doi: 10.1021/acssynbio.4c00564. PubMed (Link opens in a new window) Article (Link opens in a new window)
CIDAR MoClo Vector Expansion Kit - #1000000279
- 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
| Kanamycin | |
| Chloramphenicol | |
| Ampicillin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | DVK_p15A_AE |
|
| A / 2 | DVK_p15A_AF |
|
| A / 3 | DVK_p15A_EF |
|
| A / 4 | DVK_p15A_FG |
|
| A / 5 | DVK_p15A_GH |
|
| A / 6 | DVK_pBR322_AE |
|
| A / 7 | DVK_pBR322_AF |
|
| A / 8 | DVK_pBR322_EF |
|
| A / 9 | DVK_pBR322_FG |
|
| A / 10 | DVK_pBR322_GH |
|
| A / 11 | DVC_p15A_AE |
|
| A / 12 | DVC_p15A_AF |
|
| B / 1 | DVC_p15A_EF |
|
| B / 2 | DVC_p15A_FG |
|
| B / 3 | DVC_p15A_GH |
|
| B / 4 | DVC_pBR322_AE |
|
| B / 5 | DVC_pBR322_AF |
|
| B / 6 | DVC_pBR322_EF |
|
| B / 7 | DVC_pBR322_FG |
|
| B / 8 | DVC_pBR322_GH |
|
| B / 9 | DVA_p15A_AH |
|
| B / 10 | DVA_p15A_AG |
|
| B / 11 | DVA_p15A_AF |
|
| B / 12 | DVA_p15A_EH |
|
| C / 1 | DVA_pBR322_AH |
|
| C / 2 | DVA_pBR322_AG |
|
| C / 3 | DVA_pBR322_AF |
|
| C / 4 | DVA_pBR322_EH |
|