TABLE 3 .
Primers used for this study
| Plasmid or process and primer | Target | Template | Sequencea |
|---|---|---|---|
| pMSG360Zeo_AB | |||
| ZeoF_pMSG_F | Linear pMSG360 | pMSG360 | CGTTCTTCTTCGTCATAACTTAATG |
| ZeoR_pMSG_R | Linear pMSG360 | pMSG360 | GAAACGCCTTAAACCGGAAAATTTTC |
| Zeo_OriF | Acinetobacter ori | pABBR_MCS | tttccggtttaaggcgtttcGGATTTTAACATTTTGCGTTG |
| Zeo_OriR | Acinetobacter ori | pABBR_MCS | agttatgacgaagaagaacgGATCGTAGAAATATCTATGATTATCTTG |
| pCR-Blunt II-TOPO_AB | |||
| ZeoF_TOPO | Linear pCR-Blunt II-TOPO | pCR-Blunt II-TOPO | tcatagatatttctacgatcTTAAGGGCGAATTCTGCAG |
| ZeoR_TOPO | Linear pCR-Blunt II-TOPO | pCR-Blunt II-TOPO | aacgcaaaatgttaaaatccTCTATAGTGTCACCTAAATAGC |
| TOPOZeo_OriF | Acinetobacter ori | pABBR_MCS | GGATTTTAACATTTTGCGTTG |
| TOPOZeo_OriR | Acinetobacter ori | pABBR_MCS | GATCGTAGAAATATCTATGATTATCTTG |
| pMU125_ZeoR | |||
| ZeoR_F | Zeocin resistance cassette | pCR-Blunt II-TOPO | agcgagtcagtgagcgaggaCGTTGGCTACCCGTGATATT |
| ZeoR_R | Zeocin resistance cassette | pCR-Blunt II-TOPO | ccgcatcaggcgctcttccgGATTAGCAGAGCGAGGTATGTAG |
| pABBR_GFP | |||
| pABBR_GFP_F | eGFP | pMU125 | agcgagtcagtgagcgaggaCCCTTTCGTCTTCAAGAATTCTC |
| pABBR_GFP_R | eGFP | pMU125 | ccgcatcaggcgctcttccgTGAAGGCTCTCAAGGGCATC |
| pABBR_PuroR | |||
| PuroF1 | Linear pBacPuroR-NeoR | pBacPuroR-NeoR | GCGTCAGCGGGTGTTGGC |
| PuroR1 | Linear pBacPuroR-NeoR | pBacPuroR-NeoR | CAGTCATAGCCGAATAGCCTCTCC |
| Puro_OriF1 | Acinetobacter ori | pABBR_MCS | aggctattcggctatgactgGGATTTTAACATTTTGCGTTG |
| Puro_OriR1 | Acinetobacter ori | pABBR_MCS | ccgccaacacccgctgacgcGATCGTAGAAATATCTATGATTATCTTG |
| pBacPuroR_AB | |||
| PuroF2 | Linear pBacPuroR-NeoR | pBacPuroR-NeoR | gaggtgccgccggcttccatTCAGGCACCGGGCTTGCGGGTCA |
| PuroR2 | Linear pBacPuroR-NeoR | pBacPuroR-NeoR | aacgcagtcaggcaccgtgtATGACCGAGTACAAGCCCACGGTGC |
| Puro_OriF2 | Acinetobacter ori | pABBR_MCS | ACACGGTGCCTGACTGCG |
| Puro_OriR2 | Acinetobacter ori | pABBR_MCS | ATGGAAGCCGGCGGCACC |
| Confirmation PCR | |||
| Zeo_Confir_F1 | Zeocin resistance | pCR-Blunt II-TOPO | CGACGTGACCCTGTTCATC |
| Zeo_Confir_R1 | Zeocin resistance | pCR-Blunt II-TOPO | TCGCCGATCTCGGTCAT |
| Zeo_Confir_F2 | Kanamycin resistance | pCR-Blunt II-TOPO | CTTGTCGATCAGGATGATCTGG |
| Zeo_Confir_R2 | Kanamycin resistance | pCR-Blunt II-TOPO | CTCTTCAGCAATATCACGGGTAG |
| Puro_Confir_F1 | Puromycin resistance | pBacPuroR-NeoR | GTCACCGAGCTGCAAGAA |
| Puro_Confir_R1 | Puromycin resistance | pBacPuroR-NeoR | GGCCTTCCATCTGTTGCT |
| Puro_Confir_F2 | Amp resistance | pBacPuroR-NeoR | GCTATGTGGCGCGGTATTAT |
| Puro_Confir_R2 | Amp resistance | pBacPuroR-NeoR | CTCCGATCGTTGTCAGAAGTAAG |
| TetR_ConfirF | Tetracycline resistance | HUMC1 genomic DNA | TAAATCGCCGTGACGATCAG |
| TetR_ConfirR | Tetracycline resistance | pAT04 | GCGAGAAGCAGGCCATTAT |
a
Uppercase nucleotides represent exact matches to those in the template sequence. Lowercase nucleotides represent nucleotides in the 5′ adapter sequence needed for the Gibson assembly reaction but do not match the nucleotides in the template sequence.