P3E-EGFPpA: 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 == | ||
p3E-EGFPpA was made by PCR amplification of EGFP and an SV40 late polyadenylation signal from a pCS2+-based plasmid, using primers to add att sites, followed by a BP reaction. | |||
Note that the EGFP sequence is followed by 60 bp of polylinker sequence from pCS2+ before the stop codon. We have confirmed functionally that this cassette produces fluorescent protein in C-terminal fusion constructs. Sequencing confirms that all bases of the insert are correct. | |||
The EGFP sequence does include a start codon and a good Kozak consensus, so it is possible to use this clone to test promoters cloned into a middle clone. | |||
== Crude Map == | |||
Screenshot from ApE showing locations of components: | |||
:[[File:366 p3E-EGFPpA.PNG|thumb|none|350px|366 p3E-EGFPpA]] | |||
== | ==Download the Sequence == | ||
* '''Annotated Sequence'''(Genbank format) and '''Full-Length Sequence with individual component sequences included'''(FASTA file) | |||
: [[Media:366.p3E-EGFPpA.zip|Download p3E-EGFPpA]] | |||
Latest revision as of 20:02, 29 May 2025
Construction Details
p3E-EGFPpA was made by PCR amplification of EGFP and an SV40 late polyadenylation signal from a pCS2+-based plasmid, using primers to add att sites, followed by a BP reaction.
Note that the EGFP sequence is followed by 60 bp of polylinker sequence from pCS2+ before the stop codon. We have confirmed functionally that this cassette produces fluorescent protein in C-terminal fusion constructs. Sequencing confirms that all bases of the insert are correct.
The EGFP sequence does include a start codon and a good Kozak consensus, so it is possible to use this clone to test promoters cloned into a middle clone.
Crude Map
Screenshot from ApE showing locations of components:
366 p3E-EGFPpA
Download the Sequence
- Annotated Sequence(Genbank format) and Full-Length Sequence with individual component sequences included(FASTA file)