Table 1.

Primers Used to Develop the Probes for Southern Blot Hybridization and for PCR Analysis

Oligonucleotide name	Sequence	Size of PCR product (in bp)	Positions in RCA/SAT43 vector*
tetO-alphoid probe
tetO-R	5’-CCACTTGCACATTCTACAAATAGTGTG-3’	276	n/a
tetO-F	5’-GAGAAACTTCTTTGTGATGTTTGCATTC-3’
Vector probe 1
1F	5’-GTCGACAGCGACACACTTG-3’	200	1–200
1R	5’-AAGGGCAAGTATTGACATGTCG-3’
Vector probe 3
3F	5’-GGGCAATTTGTCACAGGG-3’	201	4243–4443
3R	5’-ATCCACTTATCCACGGGGAT-3’
Vector probe 5
5F	5’-CGCATTTTCTTGAAAGCTTTG-3’	199	9701–9900
5R	5’-TTTAAATAATCGGTGTCACTACATAAG-3’
Vector chloramphenicol gene (Cm)
Clm-F	5'-CGGAAGATCACTTCGCAGAA-3'	639	5291–5930
Clm-R	5'-CAGGGATTGGCTGAGACGAA-3'
Linker fragment homologous to RSA/SAT43 vector sequence upstream and downstream of the SpeI site
Spe/HAC-R	5’-AGGCATCTAACCTTCATGAGCA-3’	262	660–922
Spe/HAC-F	5’-CATCAGGGTGCTGGCTTTTC-3’

^*The size of the RCA/SAT vector is 10,119 bp. Position of SpeI site in the vector is 812