SBB09 Construct4

Bglbricking of M10035, PL2_Passenger_AT to Autotransporter

PCR Oso003 and Oso004 on E. coli MG1655 genomic DNA (1534 bp, EcoRI/BamHI)

Sub into pBca9145KC-Bca1144#5 (EcoRI/BamHI, 910+2974, L)

Product is pBca9145KC-M10035 {<PL2_Passenger_AT to Autotransporter!}

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Oso003 Cloning of PL2_Passenger_AT to Autotransporter  ccataGAATTCatgAGATCTacgctgacgctgaacgacag
Oso004 Cloning of PL2_Passenger_AT to Autotransporter  GCTAGggatccTCAataaccggcctgaacgcc

 You can re-use Oso002 for this construction.  Also wrong vector.  See revised below.

Bglbricking of M10035, Ag43 (autotransporter and passenger)
PCR  Oso003 and Oso002 on E. coli MG1655 genomic DNA (1600 bp, EcoRI/BamHI)
Sub into pBca9495KC-Bca1144#5    (EcoRI/BamHI, 910+2974, L)
Product is pBca9495KC-M10035    {<Ag43_long!}

 Oso003 Cloning of Ag43, long  ccataGAATTCatgAGATCTacgctgacgctgaacgacag
 Oso002 Cloning of Ag43 GCttaggatcctcagaaggtcacattcagtg