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Often, teams may seek to test multiple enzymes independently to compare their activity. To simplify the process of creating several expression vectors we used a optogentically controlled plasmid backbone wherein the red fluorescent protein mScarlet would act as our “spacer” sequence. mScarrelt dually acts as a visible selection marker to confirm that cloning was successful or not. The mScarlet ”spacer” sequence also contains a promoter, terminator, and ribosome binding site to ensure bacterial expression.There are Bbs1 cut sites flanking the Scarlet spacer region. In combination with the accompanying golden gate template (BBa0009123) teams can easily use this construct and template to assemble multiple things in only one pcr. Successful transformants are expected to appear white.  
 
Often, teams may seek to test multiple enzymes independently to compare their activity. To simplify the process of creating several expression vectors we used a optogentically controlled plasmid backbone wherein the red fluorescent protein mScarlet would act as our “spacer” sequence. mScarrelt dually acts as a visible selection marker to confirm that cloning was successful or not. The mScarlet ”spacer” sequence also contains a promoter, terminator, and ribosome binding site to ensure bacterial expression.There are Bbs1 cut sites flanking the Scarlet spacer region. In combination with the accompanying golden gate template (BBa0009123) teams can easily use this construct and template to assemble multiple things in only one pcr. Successful transformants are expected to appear white.  
  

Latest revision as of 09:00, 12 October 2023

mScarlet Optogentically controlled plasmid backbone (N-terminally His tagged)

Often, teams may seek to test multiple enzymes independently to compare their activity. To simplify the process of creating several expression vectors we used a optogentically controlled plasmid backbone wherein the red fluorescent protein mScarlet would act as our “spacer” sequence. mScarrelt dually acts as a visible selection marker to confirm that cloning was successful or not. The mScarlet ”spacer” sequence also contains a promoter, terminator, and ribosome binding site to ensure bacterial expression.There are Bbs1 cut sites flanking the Scarlet spacer region. In combination with the accompanying golden gate template (BBa0009123) teams can easily use this construct and template to assemble multiple things in only one pcr. Successful transformants are expected to appear white.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1103
    Illegal NgoMIV site found at 1235
    Illegal NgoMIV site found at 1329
    Illegal NgoMIV site found at 1622
    Illegal NgoMIV site found at 2116
    Illegal NgoMIV site found at 2134
    Illegal NgoMIV site found at 2224
    Illegal AgeI site found at 1454
    Illegal AgeI site found at 2582
    Illegal AgeI site found at 4451
  • 1000
    COMPATIBLE WITH RFC[1000]