Difference between revisions of "Part:BBa K678070"
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<partinfo>BBa_K678070 short</partinfo>
 
<partinfo>BBa_K678070 short</partinfo>
  
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p68 is a plasmid that can be used to assay promoter activity in ''Aspergillus nidulans''. It contains the'' lacZ'' gene, the TtrpC terminator, a USER cassette, an ''argB'' marker cassette for selection, an ori for propagation in E.coli, and an ''ampR'' resistance cassette. It also contains up- and down stream regions for targeting to a specific site called IS1 situated 202 bp downstream of AN6638 and 245 bp upstream of AN6639 (5). For homologous recombination to occur gene-targeting substrates have to contain these large homologous sequences around 1500 bp to ensure the targeted integration (5).
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p68 is a plasmid that can be used to assay promoter activity in ''Aspergillus nidulans''. It contains the [https://parts.igem.org/wiki/index.php?title=Part:BBa_K678026 '' lacZ''] gene, the [https://parts.igem.org/wiki/index.php?title=Part:BBa_K678036 TtrpC] terminator, a USER cassette, an ''argB'' marker cassette for selection, an ori for propagation in E.coli, and an ''ampR'' resistance cassette. It also contains up- and down stream regions for targeting to a specific site called IS1 situated 202 bp downstream of AN6638 and 245 bp upstream of AN6639 (5). For homologous recombination to occur gene-targeting substrates have to contain these large homologous sequences around 1500 bp to ensure the targeted integration (5).
  
 
To insert a promoter by USER cloning p68 should be digested with AsiSI  2-8 hours and afterwards nicked with Nb.bstI for 1-3 hours. Instructions on how to perform the USER cloning and transformation of ''A. nidulans'' can be found [http://2011.igem.org/Team:DTU-Denmark-2/Team/Protocols#USER%20cloning here].
 
To insert a promoter by USER cloning p68 should be digested with AsiSI  2-8 hours and afterwards nicked with Nb.bstI for 1-3 hours. Instructions on how to perform the USER cloning and transformation of ''A. nidulans'' can be found [http://2011.igem.org/Team:DTU-Denmark-2/Team/Protocols#USER%20cloning here].
  
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This plasmid was successfully used to characterize the ''A. nidulans'' promoters [https://parts.igem.org/wiki/index.php?title=Part:BBa_K678001 PalcA] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K678000 DMKP-P6].
 
  
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===Usage and Biology===
 
===Usage and Biology===
  
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This plasmid was successfully used to characterize the ''A. nidulans'' promoters [https://parts.igem.org/wiki/index.php?title=Part:BBa_K678001 PalcA] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K678000 DMKP-P6].
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 10:06, 20 September 2011

p68

p68 is a plasmid that can be used to assay promoter activity in Aspergillus nidulans. It contains the lacZ gene, the TtrpC terminator, a USER cassette, an argB marker cassette for selection, an ori for propagation in E.coli, and an ampR resistance cassette. It also contains up- and down stream regions for targeting to a specific site called IS1 situated 202 bp downstream of AN6638 and 245 bp upstream of AN6639 (5). For homologous recombination to occur gene-targeting substrates have to contain these large homologous sequences around 1500 bp to ensure the targeted integration (5).

To insert a promoter by USER cloning p68 should be digested with AsiSI 2-8 hours and afterwards nicked with Nb.bstI for 1-3 hours. Instructions on how to perform the USER cloning and transformation of A. nidulans can be found [http://2011.igem.org/Team:DTU-Denmark-2/Team/Protocols#USER%20cloning here].


Usage and Biology

This plasmid was successfully used to characterize the A. nidulans promoters PalcA and DMKP-P6. Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6337
    Illegal EcoRI site found at 11000
    Illegal XbaI site found at 1257
    Illegal XbaI site found at 7126
    Illegal XbaI site found at 11325
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6337
    Illegal EcoRI site found at 11000
    Illegal NotI site found at 1296
    Illegal NotI site found at 11282
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6337
    Illegal EcoRI site found at 11000
    Illegal BglII site found at 3225
    Illegal BglII site found at 8299
    Illegal BglII site found at 10490
    Illegal BamHI site found at 6407
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6337
    Illegal EcoRI site found at 11000
    Illegal XbaI site found at 1257
    Illegal XbaI site found at 7126
    Illegal XbaI site found at 11325
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6337
    Illegal EcoRI site found at 11000
    Illegal XbaI site found at 1257
    Illegal XbaI site found at 7126
    Illegal XbaI site found at 11325
    Illegal NgoMIV site found at 7043
    Illegal AgeI site found at 8847
    Illegal AgeI site found at 8877
    Illegal AgeI site found at 10254
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2122
    Illegal BsaI site found at 8680
    Illegal BsaI site found at 9852
    Illegal BsaI site found at 10446
    Illegal BsaI site found at 10642
    Illegal BsaI site found at 10813
    Illegal BsaI.rc site found at 3137
    Illegal BsaI.rc site found at 6757
    Illegal SapI.rc site found at 2793