Difference between revisions of "Part:BBa K3589106"

Revision as of 13:03, 25 October 2020

Wildtype laccase from uncultured marine bacteria for Escherichia coli

This basic part contains the coding sequence of the wildtype form of a laccase from an uncultured marine bacteria (hereafter referred to as marLac). This part is codon-optimized for Escherichia coli. Combined a promoter and a terminator, this basic part should mediate the oxidation of a wide variety of substrates including phenolic compounds and aromatic amines. This part can be cloned into standard E coli. expression vectors like the commercially available pGEX-6P-1 vector.
This laccase is cold-adapted, thermostable and shows a high tolerance for organic solvents as well as salt (Yang et. al, 2018; doi: 10.3389/fmicb.2018.02998).

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 517
Illegal PstI site found at 22
Illegal PstI site found at 52
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 517
Illegal PstI site found at 22
Illegal PstI site found at 52
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 517
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 517
Illegal PstI site found at 22
Illegal PstI site found at 52
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 517
Illegal PstI site found at 22
Illegal PstI site found at 52
Illegal NgoMIV site found at 840
Illegal AgeI site found at 93
Illegal AgeI site found at 1105
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K3589106 short</partinfo>
-This basic part contains the coding sequence of the mutant form L499F of the laccase from the ascomycete Botrytis aclada (hereafter referred to as BaLac) (B4). This part is codon-optimized for Chlamydomonas reinhardtii. Combined with the first part of BaLac (BBa_K3589105), a promoter and a terminator, this level 0 construct should mediate the oxidation of a wide variety of substrates including phenolic compounds and aromatic amines. As this part contains the introns 2 and 3 of RBCS2, it perfectly matches the part BBa_K3002027 (pAR promoter A1-B2), resulting in a high expression (Eichler-Stahlberg et al., 2009 ADD LINK). To detect the target protein, a tag of the Kaiser Collection, introduced by TU Kaiserslautern's 2019 iGEM team like, BBa_K3002017 (3xHA-tag) is recommended. This part was designed to allow synthesis due to a high GC-level. It is only functional in combination with the first part (BBa_K3589105) of the laccase. (Visit the page of the part (XXXX) to gain a deeper understanding of the functionality of BaLac.)
+This basic part contains the coding sequence of the wildtype form of a laccase from an uncultured marine bacteria (hereafter referred to as marLac). This part is codon-optimized for <I>Escherichia coli</I>. Combined a promoter and a terminator, this basic part should mediate the oxidation of a wide variety of substrates including phenolic compounds and aromatic amines. This part can be cloned into standard <I>E coli</I>. expression vectors like the commercially available pGEX-6P-1 vector.<br>
+This laccase is cold-adapted, thermostable and shows a high tolerance for organic solvents as well as salt (Yang <I> et. al</I>, 2018; doi: 10.3389/fmicb.2018.02998).
 <!-- Add more about the biology of this part here