Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Kaj Bernhardt
Date created: 2009-10-17 11:00:00
Date modified: 2015-05-08 01:11:42
pMA-BsdR-@CC2FosM
| Types | DnaRegion |
| Roles | Plasmid_Backbone plasmid_vector |
| Sequences | BBa_K260010_sequence (Version 1) |
Description
DESCRIPTION:This plasmid backbone is based on pMA from Geneart. It has BioBrick prefix and suffix flanked on both sides by 70 bp of homology arms and a blasticidin resistance cassette (BsdR) for insertion of the contained BioBrick into the proper locus of pCC2FosM ([[Part:BBa_K260000|]]) by recombineering using the HincII fragment including the BioBrick. The product of this reaction is the pCC2FosMB backbone ([[Part:BBa_K260001|]]), which then contains the same BioBrick that was inside the present backbone [[Part:BBa_K260010|]] before, with BioBrick prefix and sufix maintained intact. Initially this was done with the TT_FRT_GFP BioBrick ([[Part:BBa_K260016|]]) using [[Part:BBa_K260010|]]-[[Part:BBa_K260016|]].
The pMA-@CC2FosMB-00500bp backbone is available with [[Part:BBa_K260015|]], which is composed of promoter-FRT-dhfr. If the PvuII fragment from [[Part:BBa_K260004|]]-[[Part:BBa_K260015|]] is transferred by recombineering to the the 00500 locus of [[Part:BBa_K260001|]]-[[Part:BBa_K260016|]], the intervening sequence between the FRT sites of [[Part:BBa_K260015|]] and [[Part:BBa_K260016|]] on the [[Part:BBa_K260001|]] backbone will be 500 bp exactly.
These plasmid backbones are similar and all allow recombineering-mediated transfer of the contained BioBrick (typically [[Part:BBa_K260015|]]) to the pCC2FosMB backbone [[Part:BBa_K2600001|]]. They differ in the homology arms, which determines the locus of insertion into pCC2FosMB. The bp numbers denote final inter-FRT distance as described above:
[[Part:BBa_K260004|]]: pMA-CC2FosMB-00500bp
[[Part:BBa_K260005|]]: pMA-CC2FosMB-01000bp
[[Part:BBa_K260006|]]: pMA-CC2FosMB-02000bp
[[Part:BBa_K260007|]]: pMA-CC2FosMB-05000bp
[[Part:BBa_K260008|]]: pMA-RQ-CC2FosMB-10000bp
SOURCE:
Geneart plasmid pMA, with 2x 70 bp from pCC2FosMB [[Part:K260001|]].
DESIGN:
The 70 bp homologies were designed to delete 100 bp of [[Part:BBa_K260001|]]-[[Part:BBa_K260016|]] and put the FRT site of [[Part:BBa_K260015|]] (if used with the present backbone [[Part:BBa_K260004|]]) at a defined distance from the FRT site of [[Part:BBa_K260016|]].
Notes
The 2x 70 bp homologies were designed to delete 100 bp of pCC2FosM ([[Part:BBa_K260000]]). The insertion site on pCC2FosM was chosen to be upstream of MECP2 exon 1 but still with more than 10 kb of MECP genomic sequence upstream.Source
Geneart plasmid pMA, with 2x 70 bp from pCC2FosM ([[Part:BBa_K260000|]]).| Sequence Annotation | Location | Component / Role(s) |
| suffix ColE1 origin gb3 promoter optimised BsdR BBa_B1006 (terminator) BBa_K260000 3' homology HincII site AmpR HincII site BBa_K260000 5' homology prefix | 1,21 2149,2816 22,115 116,514 515,553 554,623 624,629 1141,2001 3000,3005 3006,3075 3076,3097 | feature/BioBrick engineered_region feature/misc sequence_feature promoter feature/promoter CDS feature/cds stem_loop feature/stem_loop sequence_feature feature/misc feature/dna sequence_feature CDS feature/cds feature/dna sequence_feature feature/misc sequence_feature feature/BioBrick engineered_region |