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Start-Stop Assembly Toolkit
(Kit # 1000000152 )

Depositing Lab:   John Heap

Start-Stop Assembly is a multi-part, modular DNA assembly method which is functionally scarless, suitable for combinatorial assembly, and ideal for metabolic engineering in diverse organisms.

This kit will be sent as bacterial glycerol stocks in 96-well plate format. Individual plasmids can be ordered from each plasmid page and will be shipped as bacterial stabs.

Add to Cart
$350 USD + shipping

Available to academics and nonprofits only.

Original Publication

Start-Stop Assembly: a functionally scarless DNA assembly framework optimised for metabolic engineering. Taylor GM, Mordaka PM, Heap JT. Nucleic Acids Research. 2019 Feb 20; doi: 10.1093/nar/gky1182. PubMed PMID: 30462270.

Description

Start-Stop Assembly is a multi-part, modular DNA assembly method which is functionally scarless, suitable for combinatorial assembly, and ideal for metabolic engineering in diverse organisms. Start-Stop Assembly provides the same key advantages as other Golden Gate-type modular assembly frameworks (efficient, unbiased, multi-part, hierarchical assembly) but also allows designs to be scarless at CDS boundaries, which are highly-sensitive sites where scars affect mRNA structure, activity of the ribosome binding site, and potentially other functional RNA features. The ‘Start-Stop Assembly Toolkit’ contains the 15 core assembly vectors used to assemble constructs of up to 15 coding sequences. It also contains the 25 promoters, RBSs, terminators and carotenoid pathway coding sequences used for assembly of metabolic pathway-encoding constructs in E. coli (described in Taylor et al. 2019). First-time users may find it helpful to assemble carotenoid pathway libraries as a positive control, because the variety of colored E. coli colonies conveniently show that combinatorial assembly is occurring (see Taylor et al. 2019 linked above for more details).

Schematic overview of Start-Stop Assembly
  • Schematic overview of Start-Stop Assembly and an example of its implementation to combinatorially assemble a carotenoid pathway.

Kit-Specific Resources

Click below to download the Start-Stop Assembly Quick-Start Guide or Start-Stop Assembly Lab Protocol:

Icon Start-Stop_Assembly_Quick_Start_Guide (57.8 KB)

Icon Start-Stop Assembly Lab Protocol (51.3 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:

“The Start-Stop Assembly Toolkit was a gift from John Heap (Addgene kit #1000000152).”

For your Reference section:

Start-Stop Assembly: a functionally scarless DNA assembly framework optimised for metabolic engineering. Taylor GM, Mordaka PM, Heap JT. Nucleic Acids Research. (2019) doi: 10.1093/nar/gky1182. PubMed PMID: 30462270.

Start-Stop Assembly Toolkit - #1000000152

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
Tetracycline
Kanamycin
Chloramphenicol

Inventory

Well Plasmid Resistance
A / 1 pStA0::J23100
Ampicillin
A / 2 pStA0::J23102
Ampicillin
A / 3 pStA0::J23107
Ampicillin
A / 4 pStA0::J23113
Ampicillin
A / 5 pStA0::J23116
Ampicillin
A / 6 pStA0::J23118
Ampicillin
A / 7 pStA0::RBSc13
Ampicillin
A / 8 pStA0::RBSc33
Ampicillin
A / 9 pStA0::RBSc44
Ampicillin
A / 10 pStA0::RBSc58
Ampicillin
A / 11 pSTA0::RBSc36
Ampicillin
A / 12 pStA0::RBSc42
Ampicillin
B / 1 pStA0::L3S2P55
Ampicillin
B / 2 pStA0::L3S2P21
Ampicillin
B / 3 pStA0::ECK120033737
Ampicillin
B / 4 pStA0::ECK120019600
Ampicillin
B / 5 pStA0::crtI
Ampicillin
B / 6 pStA0::crtB
Ampicillin
B / 7 pStA0::crtE
Ampicillin
B / 8 pStA0::dxs
Ampicillin
B / 9 pStA0::crtW
Ampicillin
B / 10 pStA0::crtZ
Ampicillin
B / 11 pStA0::lcyB
Ampicillin
B / 12 pStA0::idi
Ampicillin
C / 1 pStA0::eyfp
Ampicillin
C / 2 pStA0
Ampicillin
C / 3 pStA1AZ
Tetracycline
C / 4 pStA1AB
Tetracycline
C / 5 pStA1BZ
Tetracycline
C / 6 pStA1BC
Tetracycline
C / 7 pStA1CZ
Tetracycline
C / 8 pStA1CD
Tetracycline
C / 9 pStA1DZ
Tetracycline
C / 10 pStA1DE
Tetracycline
C / 11 pStA1EZ
Tetracycline
C / 12 pStA212
Kanamycin
D / 1 pStA223
Kanamycin
D / 2 pStA234
Kanamycin
D / 3 pStA313
Chloramphenicol
D / 4 pStA314
Chloramphenicol
Data calculated @ 2019-11-12

Kit Plate Map - #1000000152

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