BioBrick integrative base vector for S. cerevisiae
Susanna Zucca, Lorenzo Pasotti, Federica Sampietro, Paolo Magni
NheI restriction enzyme site (XbaI, SpeI compatible overhang)
NheI recognition site. Digestion with NheI generates a compatible cohesive end to XbaI and SpeI.
[LoxP-KanMX-LoxP] excisable marker for yeast
This cassette is to be used in yeast. It is composed by the kanMX gene from the E. coli transposon Tn903 that confers the resistance to the aminoglycoside antibiotic G418 to yeast (commonly used at 200 ug/ml working concentration). This gene is expressed by the TEF2 promoter/RBS and terminated by the TEF2 transcriptional terminator from Ashbya gossypii filamentous fungus. The cassette is flanked by two loxP sites and for this reason it can be excised with Cre/loxP mediated marker removal recombination procedure. ADVANTAGES IN GENOME ENGINEERING APPLICATIONS: This part can be used to select positive integrants in the yeast genome and then it can be eliminated through Cre/loxP procedure. This enables the engineering of yeast genome in multiple steps, always using the same dominant selection marker.
The original sequence of loxP-KanMX-loxP from pUG6 vector has two NsiI restriction sites in kanR(Tn903) coding sequence and they have been removed in this part by changing codons. The TEF2 promoter has a XbaI and a PstI restriction sites, which have not been removed to avoid the change of the promoter strength. For this reason, this part is not fully compatible with RFC10 BioBrick Standard Assembly, but it can be assembled by using EcoRI-SpeI enzymes.
Lorenzo Pasotti, Giacomo Zambianchi, Federica Sampietro, Paolo Magni
This is a part consisting of only a single nucleotide ... T. It can act as a spacer between restriction sites and parts.
This part is the standard BioBricks prefix.
EcoRI (<partinfo>BBa_M31983</partinfo>) and MfeI (<partinfo>BBa_B0047</partinfo>) have a compatible overhang (AATT). The EcoRI/MfeI mixed site is GAATTG.
Susanna Zucca, Federica Sampietro, Paolo Magni
NsiI (<partinfo>BBa_B0046</partinfo>) and PstI have a compatible overhang (TGCA). The NsiI/PstI mixed site is ATGCAG.
This site can also be cut with PstI restriction enzyme.
This part is identical to pSB1A2, but it does not have the Prefix and Suffix. It can be useful to specify the sequence of new vectors constructed with BioBrick parts, such as <partinfo>BBa_K300001</partinfo>.
No changes have been made to <partinfo>pSB1A2</partinfo> sequence.
Homologous region in S288C yeast genome (GAL10 ORF)
Homologous region in S288C yeast genome (GAL1 ORF)
AvrII restriction enzyme site (XbaI, SpeI compatible overhang)
AvrII recognition site. Digestion with AvrII generates a compatible cohesive end to XbaI and SpeI.
Conversion of the iGEM parts registry to SBOL2.1