Supplementary Figure 1. Inverse-PCR Cloning of Pstark5.5

Supplementary Figure 1. Inverse-PCR cloning of pStark5.5

The ~5.5 kb band produced by hybridizing pBGBM18.2 to a Southern blot digested with BamHI was cloned using a PCR strategy. 1) Outward facing primers were designed from the pBGBM18.2 sequence. 2) Genomic DNA was digested with BamHI, self-ligated and 3) used as a template for a PCR reaction producing 5.5 kb and ~11 kb products. 4) The PCR products were cloned using a TA-cloning kit.

Supplementary Figure 2. CentC is not present in fosmids that contain pStark-1.

90 fosmids putatively identified as containing pStark-1 were dotted onto a nylon membrane and hybridized with CentC (top panel). The three spots with white stars are controls containing PCR amplified CentC, cloned CentC, or genomic DNA. The blot was stripped and hybridized with pStark-1 (lower panel). No fosmids were identified that contain both CentC and pStark-1.

Supplementary Table 1. List of primer sequences

Primer name / Sequence
StarkF / ttg atg gac gac taa gat t
StarkR / ttc gag aac cac taa aaa
StarkF_RC / aat ctt agt cgt cca tca a
StarkR_RC / ttt tta gtg gtt ctc gaa
2NB1.RC / atc aag aac tca acg gtc ac
2NB1 / gtg acc gtt gag ttc ttg at
2NB2 / tct tga tgg acg act aag att
2NB2.RC / aat ctt agt cgt cca tca aga
2NB3.RC / ttt tta gtg gtt ctc gaa cct
2NB3 / agg ttc gag aac cac taa aaa
2NB4.RC / aac caa ttt gga ttc atg cct
2NB4 / ggc atg aat cca aat tgg tt
46F1R / cta agg caa ata gat aag ggc
46F1F / gcc ctt atc tat ttg cct tag
46F2R / cct ttc gtg agg cct tct t
46F2F / aag aag gcc tca cga aag g
46F3R / gat aac gtt tga att cag tga g
46F3F / ctc act gaa ttc aaa cgt tat c
46F4R / ggg tcc gac tgc aca ctc
46F4F / gag tgt gca gtc gga ccc
46F5F / gtt ctg ctt cgg aac cat ct
46F5R / aga tgg ttc cga agc aga ac
46F6F / gag atc cac agg tat caa ctt
46F6R / aag ttg ata cct gtg gat ctc
46F7R / ctt gaa aca acg atc tcg tag
46F7F / cta cga gat cgt tgt ttc aag
46R1F / gac aca ggg gtc tta gtg a
46R1R / cca cta aga ccc ctg tgt c
46R1R / tca cta aga ccc ctg tgt c
46R2F / cga tac aag gat gag atg gg
46R2R / ccc atc tca tcc ttg tat cg
46R3F / gct gag acg tga aga tgg g
46R3R / ccc atc ttc acg tct cag c
46R4F / gac tga ttg gat gcg tgt gca
46R4R / tgc aca cgc atc caa tca gtc
46R5F / tac cgg cta tgt gtg cgt g
46R5R / cac gca cac ata gcc ggt a
46R6F / ggt att caa att cga att cac ta
46R6R / tag tga att cga att tga ata cc
46R7F / tcc aaa ttt aaa ctt gtc tca ca
46R7R / tgt gag aca agt tta aat ttg ga

Supplementary Table 2: Primer pairs producing RT-PCR products:

Primer 1, Primer 2
2NB1, 2NB4
46F1F, 46F2R
46F1F, 46F3R
46F3F, 46R4R
46R3F, 46R7R
46R3R, 2NB4
2NB1, 46F3R
46F6F, 46R7F

1