Table 2 Plasmids used for integrative vector construction

pHC-attP-CS

pMK-RQ(BBa_K300983)^a

DNA fragment including FRT-NheI-attP-NheI-CS(B0033)-FRT, flanked by AvrII sites, in high copy vector

pHC-CmR

pSB1A1(BBa_P1004)^b

Chloramphenicol resistance gene with promoter in high copy vector

pHC-ter

pSB1AK3(BBa_B0015)^b

Double terminator in high copy vector

pHC-R6K

pSB1A2(BBa_J61001)^b

R6K conditional replication origin in high copy vector

pHC-CmRter

pSB1AK3(BBa_K300032)^c

Chloramphenicol resistance cassette in high copy vector

pHC-PR-RFP

pSB1A2(BBa_I763007)^b

RFP constitutive expression cassette, driven by the PR promoter in high copy vector

pHC-CmRter-R6K

pSB1A2(BBa_K300008)^c

Chloramphenicol resistance cassette with R6K origin downstream in high copy vector

pBBintФ-B0033

BBa_K300982^c

Integrative base vector, targeting the Ф80 locus, with RBS BBa_B0033 as insert

pBBintФ-RFP

BBa_K300000(BBa_I763007)^c

Integrative base vector, targeting the Ф80 locus, with RFP constitutive expression cassette as insert

pHC-aspA

pSB2K3(BBa_C0083)^b

aspA coding sequence in high copy vector

pBBintAsp-RFP

BBa_J107058(BBa_I763007)^c

Integrative vector, specialized to target the aspA locus, with RFP constitutive expression cassette as insert

pInt80-649

BBa_J72008^d

Ampicillin-resistant low copy helper plasmid, with temperature-sensitive pSC101 origin, containing a constitutively expressed pir-116 gene and the site-specific recombination machinery of the Ф80 phage under the control of a heat-sensitive promoter.

pCP20

not in the Registry^e

Ampicillin- and chloramphenicol-resistant low copy helper plasmid, with temperature-sensitive pSC101 origin, containing the yeast Flp recombinase under the control of a heat-sensitive promoter.