Name Sequence (5΄-3΄)a Sequence (5΄-3΄)a Purpose
Sph-E Nco-E Ack-Gen-F Ack-E-R Ack-F Gen-F Ack-catF Ack-catR TetA-F Ack-R EG-pkd-E EG-pkd-G Pkd-cx Gen-R PigA-EGF PigA-EGR Pig-520F pigA-ssF PigA-T7-R1 AraB-T7-F Pig-1690R T7-F Ack-EGSD Arab-EGSDF Arab-EGR Arab-EGF Arab-F Arab-R E-R Hyp-EGSDF Hyp-EGSDR Mar-EGSDF Mar-EGSDR Rcs-EGSDF Rcs-EGSDR Sph-E Nco-E Ack-Gen-F Ack-E-R Ack-F Gen-F Ack-catF Ack-catR TetA-F Ack-R EG-pkd-E EG-pkd-G Pkd-cx Gen-R PigA-EGF PigA-EGR Pig-520F pigA-ssF PigA-T7-R1 AraB-T7-F Pig-1690R T7-F Ack-EGSD Arab-EGSDF Arab-EGR Arab-EGF Arab-F Arab-R E-R Hyp-EGSDF Hyp-EGSDR Mar-EGSDF Mar-EGSDR Rcs-EGSDF Rcs-EGSDR CCAGCATGCATGGTACGCTGGACTTTGT ACTCCATGGTCACTCCTTCCGCACGTAA CTGACGTTTTTTTAGCCACGTATCAATTATAGGTACTTCCTTAGGTGGCGGTACTTGGG ATTAATACTGAAACTGAGATCAAGCAAAAGCATTCACTAATTGACATAAGCCTGTTCGG TGCAGTGCATGATGTTAATC TTGACATAAGCCTGTTCGG TGACGTTTTTTTAGCCACGTATCAATTATAGGTACTTCCAGCTACCAGATACCATCGTC TTACAAAACAGCACCGCCAGCTGAGCTGGCGGTGTGAAATAAGAGCGCATGATACGGAC CAAGAGGGTCATTATATTTCG GGGATCAGCATAATAATACG GTTTTTTTGGGAATTCGAGCTCTAAGGAGGTTATAAAAAATGGTACGCTGGACTTTGT AAGTGATTGCGCCTACCCGGATATTATCGTGAGGATGCGTTAGGTGGCGGTACTTGGG GCTCTTCTCGCTAACCAAAC TTAGGTGGCGGTACTTGGG ACTGATCGCGGCGAATGCGGCGACGGAGCCGGACGCCGGTTAGGTGGCGGTACTTGGG CAGGGTTGAGCGGTTGATTTCATGGCATAAATATCGGAATTATGACAACTTGACGGCTA AACGAATCGTCTTGCTGGTT ACTGATCGCGGCGAATGCGGCGACGGAGCCGGACGCCGG//TTCCGATATTTATGCCATGAAATCAACCGCTCAACCCTG ACTGATCGCGGCGAATGCGGCGACGGAGCCGGACGCCGGTACGTTGGCGACCGCTAGG CTGTTTCTCCATACCCGTTTTTTTGGATGGAGTGAAACGATGAACACGATTAACATCG CCAGATCATTGAATCGTGGA ATGAACACGATTAACATCG AATACTGAAACTGAGATCAAGCAAAAGCATTCACTAAGAAGGAGATATACCATGGTACGCTGGACTTTG CTGTTTCTCCATACCCGTTTTTTTGGATGGAGTGAAACGATGTGCCAATCGCGGGGGGT CACACTTCATAATTATCAAAAATCGTCATTATCGTGTCCTTAGGTGGCGGTACTTGGGT CTGTTTCTCCATACCCGTTTTTTTGGATGGAGTGAAACGATGGTACGCTGGACTTTGTG TCCATAAGATTAGCGGATC ATGTTGGGTGACCTGACGCA TCACTCCTTCCGCACGTAAT AGATGCTAGTGATATGCTTATTAACATTTCTGAATACTTTTATGACAACTTGACGGCTA ATAGTAATCTCCCATTTCACTTTCGCCATCATAACCAACTTAGGTGGCGGTACTTGGGT AGAGTTATCTTGGAATAAGTTAGGGCGAGGCGGAGAAATTTATGACAACTTGACGGCTA ATAATCTGTACCGGTTGAAAACGTGGGCAAAAACCGGTGTTAGGTGGCGGTACTTGGGT ATATTACCGCAGTAAAACGGATTTAAGTGGGGGTGCGATTTATGACAACTTGACGGCTA TAACGGCTGGTAAAACAGAGCGGGGGCGTTCTCTGTGGTTTAGGTGGCGGTACTTGGGT Construction of plasmid pBBR1-E Replacement of kil by E-GmR Identification of strain CWE-1 Replacement of ack by partial fragment of catenin gene Identification of strain CWE-2 Insertion of E-GmR after promoter PBAD in plasmid pKD46 Identification of plasmid pKD-EG Insertion of PBAD-E-GmR double selection cassette at pigA locus Paired with Gen-F to identify strain GY4 Reparation of PBAD-E-GmR insertion mutation using ssDNA Replacement of PBAD-E-GmR by partial fragment of T7 RNA polymerase gene Paired with Pig-520F to identify PBAD-E-GmR deletion by ssDNA Paired with Pig-520F to identify GY4-1 Paired with Ack-Gen-F to replace kil at ack locus Replacement of araB by PBAD-kil-sd-E- GmR cassette Paired with Arab-EGR to Replacement of araB by PBAD-E- GmR cassette Identification of gene Replacement at araB locus Paired with Arab-F to identify MG-4A/4B Replacement of hyp-1 by PBAD-kil-sd-E- GmR cassette Replacement of marR-1 by PBAD-kil-sd-E- GmR cassette Replacement of rcsA by PBAD-kil-sd-E- GmR cassette