Figure 1.

Design and assembly of multi-part constructs from multiplexed, retrievable pools. The input to the pool extraction algorithm is a library of constructs that can be designed computationally or by hand. Dashed lines in the constructs indicate the absence of a gene. The extraction algorithm designs pools of composite parts that encompass the entire design space (combinations of parts). The parts for assembling multi-gene constructs after retrieval are included in the pool design; these can include Type IIS recognition sites and 4 bp linkers (diamonds). Barcode symbol represents the random 20 bp barcodes used for retrieval. Red X's indicate mutations in the composite part construct. Green ovals represent a constructed composite part whose sequenced reads perfectly match a designed composite part in the pool. The pools are colored to correspond to the gene from the desired constructs contained in the composite parts. The numbers below each pool are examples of the number of sequence-verified unique composite parts contained (out of 5000 designs in each pool).