P5E-bactin2: Difference between revisions
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Hggenusrwki (talk | contribs) Created page with "== Construction Details == This template is used to display DNA sequence information in a standardized format. == Crude Map == photo showing locations of components == Sequence == * Annotated Sequence: (Genbank format) link * Full-Length Sequence with individual component sequences included: (FASTA file) link == Reference == This template is used to display DNA sequence information in a standardized format." |
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== Construction Details == | == Construction Details == | ||
p5E-MCS was made by PCR of the pBSII SK+ multiple cloning site, from T7 to T3 primers, followed by a BP reaction with pDONR P4-P1R. Sequencing confirms that all bases of the insert are correct. | |||
M13F, M13R, and T3 should all work as sequencing primers. | |||
Unfortunately, the backbone of P4-P1R contains recognition sites for several potentially useful restriction enzymes. The restriction sites from the Bluescript MCS that remain unique in p5E-MCS are: | |||
''Asp718-KpnI-DraII-XhoI-SalI-Bsp106-HindIII-SmaI-BamHI-SpeI-SacII-BstXI'' | |||
== Crude Map == | == Crude Map == | ||
Screenshot from Sequencher showing locations of components: | |||
P5E-MCS.png | |||
== Sequence == | ==Download the Sequence == | ||
* Annotated Sequence | * Annotated Sequence (Genbank format) and Full-Length Sequence with individual component sequences included (FASTA file) | ||
: [[Media:233._pME-nlsmCherry.zip|Download p5E-MCS]] | |||
== Reference == | == Reference == | ||
This template is used to display DNA sequence information in a standardized format. | This template is used to display DNA sequence information in a standardized format. | ||
Revision as of 15:57, 11 March 2025
Construction Details
p5E-MCS was made by PCR of the pBSII SK+ multiple cloning site, from T7 to T3 primers, followed by a BP reaction with pDONR P4-P1R. Sequencing confirms that all bases of the insert are correct.
M13F, M13R, and T3 should all work as sequencing primers.
Unfortunately, the backbone of P4-P1R contains recognition sites for several potentially useful restriction enzymes. The restriction sites from the Bluescript MCS that remain unique in p5E-MCS are:
Asp718-KpnI-DraII-XhoI-SalI-Bsp106-HindIII-SmaI-BamHI-SpeI-SacII-BstXI
Crude Map
Screenshot from Sequencher showing locations of components: P5E-MCS.png
Download the Sequence
- Annotated Sequence (Genbank format) and Full-Length Sequence with individual component sequences included (FASTA file)
Reference
This template is used to display DNA sequence information in a standardized format.