Difference between revisions of "Part:BBa K4806005"
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<h2>Sequence and Features</h2>
 
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<h2>Results</h2>
 
<h2>Results</h2>

Revision as of 11:52, 20 September 2023


CYP81A10V7 gene for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the coding sequence of CYP81A10V7 (B3-B4). This part is codon-optimized for Chlamydomonas reinhardtii and was built as part of the CYPurify Collection. In combination with a promoter like AβSAP(i) (BBa_K4806013) and a terminator like tRPL23 (BBa_K3002006)*, this level 0 part leads to expression and potential detoxification of specific chemicals (Ohkawa & Inui, 2015). To detect the target protein a tag like HA-tag (BBa_K3002017)* is recommended.


Constructs

Fig.1 Construct design
We designed 3 level 2 constructs containing CYP81A10V7 using the modular cloning system (MoClo).


Here are the links to the built constructs:

  • 1. CYP2D6 gene with HA-tag for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806219)
  • 2. CYP2D6 gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806220)
  • 3. CYP2D6 gene with mNeonGreen for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806221)

These constructs were transformed into Chlamydomonas reinhardtii. Besides the CYP81A10V7 coding sequence the constructs contain the AβSAP(i)-promotor (BBa_K4806013), the HA-tag (BBa_K3002017)* or mNeonGreen (BBa_K4806006) for detection or mStop (BBa_K4806009) and the tRPL23-terminator (BBa_K3002006)*. The resistance cassette for spectinomycin is already built in the level 2 vector pMBS807 we are using. The usage of this vector allows the direct assembly of level 0 parts to level 2 constructs, facilitating the cloning time (Niemeyer & Schroda, 2022).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1685
    Illegal PstI site found at 1827
    Illegal PstI site found at 2449
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1685
    Illegal PstI site found at 1827
    Illegal PstI site found at 2449
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1685
    Illegal PstI site found at 1827
    Illegal PstI site found at 2449
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1685
    Illegal PstI site found at 1827
    Illegal PstI site found at 2449
    Illegal NgoMIV site found at 1532
  • 1000
    COMPATIBLE WITH RFC[1000]


Results