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Thermo Scientific pTZ19R/U are small phagemids 2862 bp in length, constructed by inserting the DNA of phage f1 intergenic region (IG) into pUC19 and the T7 promoter sequence near to the MCS of pUC19. The phagemids differ in the orientation of the cloned f1 IG region. They are designed for DNA cloning, dideoxy DNA sequencing, in vitro mutagenesis, and in vitro transcription in one system. A host strain harboring these phagemids will produce single-stranded DNA if superinfected with the helper phage M13K07.
pTZ19R DNA map. Download pTZ19R sequence in FASTA, GenBank, or EMBL format.
For DNA sequence, sequence analysis and map creation, see the free REviewer online tool.
Synthesis of single-stranded (plus) DNA requires phage-encoded gene II, X, and V proteins. It is initiated at ori (+) and proceeds in the direction indicated. The conversion of plus DNA strands to double-stranded DNA does not require any of the phage genes. DNA synthesis is initiated by a 30-nucleotide RNA primer synthesized by the host’s RNA polymerase and starting at ori (-).
The map shows enzymes that cut pTZ19R DNA once. Enzymes produced by Thermo Scientific are shown in orange. The coordinates refer to the position of the first nucleotide in each recognition sequence.
The exact positions of the genetic elements are shown on the map (termination codons included). The bla gene nucleotides 2732-2664 (complementary strand) code for a signal peptide. The LacZ polypeptide corresponding to wt beta-galactosidase and essential for blue/white screening ends at nt position 458 (complementary strand). The remaining amino acids in the LacZ reading frame are encoded by f1 DNA. The indicated rep region is sufficient to promote replication. DNA replication initiates in the complementary DNA strand at position 1112 (± 1) and proceeds in the direction indicated. Phagemids and plasmids carrying the pMB1 and ColE1 replicons are incompatible with one another, but are fully compatible with those carrying p15A replicon (pACYC177, pACYC184). pMB1-derived plasmids can be amplified using chloramphenicol.
D. A. Mead et al., Single-stranded DNA ‘blue’ T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng. 1, 67-74 (1986).