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Biological and Medical Sciences > Classification by Description > Reagents, Kits and Assays > DNA and RNA > Cloning

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Branch: Sci-Wiki Type: General Knowledge Intention: For Publication Read/Write Permissions: Open Access

Authors : 16.09.2010 - John Stewart    13.09.2010 - Christopher Dyer  -  Sci-Mate

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The aat sites within the Gateway system have been modified to eliminate stop codons, increase specificity, minimize secondary structure formation, and increase efficiency of the recombination reaction, see Gateway Cloning. 

Table 1: Gateway restriction and ligation sites (bold bases represent 5'-3' overhangs)

Gateway aatB1 sequence:	5'-ACA-AGT-TTG-TAC-AAA-AAA-GCA-GGC-T-3'
Gateway aatB2 sequence:	5'-ACC-ACT-TTG-TAC-AAG-AAA-GCT-GGG-T-3'
Gateway aatP1 sequence:	5'-CCA-ACT-TTG-TAC-AAA-AAA-GCT-GAA-C-3'
Gateway aatP2 sequence:	5'-CCA-ACT-TTG-TAC-AAG-AAA-GCT-GAA-C-3'
Gateway aatL1 sequence:	5'-CCA-ACT-TTG-TAC-AAA-AAA-GCA-GGC-T-3'
Gateway aatL2 sequence:	5'-CCA-ACT-TTG-TAC-AAG-AAA-GCT-GGG-T-3'
Gateway aatR1 sequence:	5'-ACA-AGT-TTG-TAC-AAA-AAA-GCT-GAA-C-3'
Gateway aatR2 sequence:	5'-ACA-ACT-TTG-TAC-AAG-AAA-GCT-GAA-C-3'

Table 2: If presented in the following order, one can more easily see how the B1 and P1 sites recombine to form L1 and R1 sites; and how B2 and P2 sites recombine to form L2 and R2 sites. 

Gateway aatB1 sequence:	5'-ACA-AGT-TTG-TAC-AAA-AAA-GCA-GGC-T-3'
Gateway aatP1 sequence:	5'-CCA-ACT-TTG-TAC-AAA-AAA-GCT-GAA-C-3'
Gateway aatL1 sequence:	5'-CCA-ACT-TTG-TAC-AAA-AAA-GCA-GGC-T-3'
Gateway aatR1 sequence:	5'-ACA-AGT-TTG-TAC-AAA-AAA-GCT-GAA-C-3'
Gateway aatB2 sequence:	5'-ACC-ACT-TTG-TAC-AAG-AAA-GCT-GGG-T-3'
Gateway aatP2 sequence:	5'-CCA-ACT-TTG-TAC-AAG-AAA-GCT-GAA-C-3'
Gateway aatL2 sequence:	5'-CCA-ACT-TTG-TAC-AAG-AAA-GCT-GGG-T-3'
Gateway aatR2 sequence:	5'-ACA-ACT-TTG-TAC-AAG-AAA-GCT-GAA-C-3'

See also: Designing Gateway PCR Primers