Table 2.
Plasmids used for integrative vector construction
| Name | BioBrick™ code | Description |
|---|---|---|
| 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_K300982c |
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_J72008d |
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 Registrye | 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. |
The BioBrick™ code of the plasmids include the vector name out of brackets and insert name in brackets.
a: obtained from Mr. Gene DNA synthesis service. pMK-RQ is a kanamycin-resistant high copy number delivery vector from Mr. Gene.
b: obtained from DNA Distributions of the MIT Registry of Standard Biological Parts.
c: constructed in this study.
d: gift from the Dr. J.C. Anderson lab (UC Berkeley).
e: obtained from CGSC (plasmid purified from recombinant strain CGSC#7629).