Part:BBa_K3376013
BioBrick / pDL278
pDL278, a E. coli/Streptococcus shuttle vector
pDL278 is a shuttle vector between E. coli and Gram-positive bacteria esp. for Streptococcus spp., which was created by Donald J. LeBlanc, et al. in 1992. It could be applied for cloning vectors in E. coli and transforming S. mutans with a spectinomycin resistance cassette and the origin of replication of pBR322 for E. coli and ori (+) for Gram(+) bacteria, respectively. Ori (+) is from S. aureus with an ORF encoding an undefined protein possibly for plasmid replication (Fig. 1, A). We got the plasmid from the lab of Dr. Yuqing Li at Sichuan University in China. Firstly, we made the pDL278 compatible to BioBrick assembly system (i.e., EcoRI-XbaI-insert-SpeI-PstI). We amplified the backbone by PCR and did 2 rounds of site-directed mutagenesis with the primers listed in Fig. 1, C. The resulting plasmid was checked by restriction enzymes (Fig. 20, B) and further confirmed by sequencing.
Transformation
The ldhp-GFP-Tr/pSB1C3 [BBa_K3376002] was transformed into E. coli Nissle strain. The high expression of green fluorescent protein was observed under a blue led light.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3022
Illegal SapI site found at 1991
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