Team:Bielefeld-CeBiTec/Basic Part

Basic Parts
The broad range nucleotide transporter PtNTT2 from the algae Phaeodactylum tricornutum facilitates the counter exchange of nucleotide triphosphates (NTPs) and nucleoside diphosphates (NDPs), importing NTPs into the plastid. Expressed in E. coli, it allows the uptake of various nucleotides from the cultivation media. This function is essential for the development of a semisynthetic organism, which uses externally provided unnatural nucleotides for replication of its semisynthetic DNA. For our project, we characterized the structure, kinetics, dynamics and subcellular localization of PtNTT2 and different PtNTT2 variants. Therefore, we applied bioinformatic prediction tools, confocal laser scanning microscopy, SDS-PAGE, Western Blot as well as HPLC-MS. All PtNTT2 variants were also cultivated to investigate the effect of PtNTT2 expression on the growth rate. In the future, advanced endosymbiotic systems or novel biosafety mechanisms can be developed based on this part.
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Name Type Description Designer Length
BBa_K2201004 Coding Nucleotide Transporter PtNTT2 from Phaeodactylum tricornutum Christopher M. Whitford 1728
BBa_K2201000 Coding Nucleotide Transporter PtNTT2 with lacUV5 Promoter and RBS Christopher M. Whitford 1777
BBa_K2201001 Coding Truncated version PtNTT2(66-575) with lacUV5 and RBS Christopher M. Whitford 1585
BBa_K2201002 Coding PtNTT2 tagged with GFP and a cMyc linker Christopher M. Whitford 2518
BBa_K2201003 Coding PtNTT2(66-575) tagged with GFP and a cMyc linker Christopher M. Whitford 2326
BBa_K2201005 Coding Truncated version PtNTT2(31-575) with lacUV5 and RBS Christopher M. Whitford 1690
BBa_K2201006 Coding PtNTT2 with pelB Signal Peptide, lacUV5 Promoter and RBS Christopher M. Whitford 1753
BBa_K2201007 Coding PtNTT2 with TAT Signal Peptide, lacUV5 Promoter and RBS Christopher M. Whitford 1777
BBa_K2201008 Regulatory pSC101 temperature sensitive origin of replication Christopher M. Whitford 1221
BBa_K2201009 DNA Flanking sequences for deletion of codA Christopher M. Whitford 1369
BBa_K2201010 Coding Construct comprised of PtNTT2, lacO and Cas9 Christopher M. Whitford 6155
BBa_K2201011 Coding PtNTT2(31-575) Tagged with GFP and a cMyc Linker Christopher M. Whitford 2431
BBa_K2201012 Coding PtNTT2 with pelB Signal Peptide Tagged with GFP and a cMyc Linker Christopher M. Whitford 2494
BBa_K2201013 Coding PtNTT2 with TAT signal peptide tagged with GFP and a cMyc linker Christopher M. Whitford 2518
BBa_K2201017 Coding Construct for the integration of unnatural base pairs Christopher M. Whitford 2167
BBa_K2201020 Regulatory lacO_tight1 from E. coli BL21(DE3) and optimized Lennard Karsten 118
BBa_K2201021 DNA Left flanking sequence for HDR-mediated genomic integration in E. coli BL21(DE3) Lennard Karsten 985
BBa_K2201022 DNA right flanking sequence for genomic integration into E. coli BL21(DE3) Lennard Karsten 986
BBa_K2201023 Coding cas9 from S. pyogenes + rho-independent trp RNA-terminator Lennard Karsten 4147
BBa_K2201026 Coding Plasmid for the integration of unnatural base pairs Christopher M. Whitford 904
BBa_K2201027 Coding PtNTT2(31-575)-lacO-Cas9 construct Christopher M. Whitford 6068
BBa_K2201029 Regulatory lacO_tight2 from E. coli BL21(DE3) and optimized Lennard Karsten 32
BBa_K2201030 Coding cas9 from Streptococcus pyogenes Lennard Karsten 4107
BBa_K2201031 Terminator trp RNA-Terminator rho-independent Lennard Karsten 32
BBa_K2201050 Measurement Cleavage of Carotenoids Michelle Liebers 1887
BBa_K2201060 Coding GMPS iso1 Markus Haak 1605
BBa_K2201061 Coding GMPS iso2 Markus Haak 1605
BBa_K2201062 Coding CDA Markus Haak 942
BBa_K2201100 DNA random spacer sequence Markus Haak 25
BBa_K2201101 DNA random spacer sequence Markus Haak 25
BBa_K2201102 DNA random spacer sequence Markus Haak 25
BBa_K2201103 DNA random spacer sequence Markus Haak 25
BBa_K2201104 DNA random spacer sequence Markus Haak 25
BBa_K2201105 Regulatory Fis TFBS + proK promotor Markus Haak 93
BBa_K2201106 DNA gRNA scaffold Markus Haak 76
BBa_K2201107 Terminator proK terminator Markus Haak 35
BBa_K2201108 DNA spacer Markus Haak 2
BBa_K2201109 DNA spacer Markus Haak 2
BBa_K2201110 DNA spacer Markus Haak 7
BBa_K2201111 RNA crRNA R-target mutA Markus Haak 18
BBa_K2201112 RNA crRNA R-target mutG Markus Haak 18
BBa_K2201113 RNA crRNA R-target mut∆ Markus Haak 18
BBa_K2201114 RNA crRNA R-target mutT Markus Haak 18
BBa_K2201115 RNA crRNA R-target mutC Markus Haak 18
BBa_K2201116 RNA crRNA MAX-target mutA Markus Haak 18
BBa_K2201117 RNA crRNA MAX-target mutG Markus Haak 18
BBa_K2201118RNA crRNA MAX-target mut∆ Markus Haak 18
BBa_K2201119 RNA crRNA MAX-target mutT Markus Haak 18
BBa_K2201120 RNA crRNA MAX-target mutC Markus Haak 18
BBa_K2201180 Intermediate lacUV5 and RBS Christopher M. Whitford 49
BBa_K2201181 Coding Myc (human c-Myc oncogene) epitope tag Christopher M. Whitford 30
BBa_K2201200 Coding Tyrosyl tRNA/aminoacyl-synthetase for the incorporation of 2-nitrophenylalanine Yannic Kerkhoff 1738
BBa_K2201205 Coding Cysteinyl-lysinyl-tRNA Synthetase Daniel Bergen 1474
BBa_K2201206 RNA pyrrolysinyl-tRNA Daniel Bergen 378
BBa_K2201207 Coding Expression plasmid of an aminoacyl-tRNA synthetase for AzoPhe Yannic Kerkhoff 1749
BBa_K2201220 Coding CDS of GFP-streptavidin fusion protein with medium gly-gly-ser linker Yannic Kerkhoff 1131
BBa_K2201221 Coding CDS of GFP-streptavidin fusion protein with medium gly-gly-ser linker containing an amber-codon Yannic Kerkhoff 1131
BBa_K2201230 Coding NM-domain of Sup35 with an amber codon at position 21 Svenja Vinke 753
BBa_K2201231 Coding NM-domain of Sup35 with an amber codon at position 21 and histidine tag. Svenja Vinke 786
BBa_K2201241 Generator CrtEBI under constitutive promoter with an amber codon at amino acid position 318 of crtI Svenja Vinke 3448
BBa_K2201242 Generator CrtEBI under constitutive promoter with an amber codon at amino acid position 353 of crtI Svenja Vinke 3448
BBa_K2201260Coding Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and mRFP fusion protein Svenja Vinke 2503
BBa_K2201261 Coding Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with an amber stop codon at amino acid po Svenja Vinke 1791
BBa_K2201262 Coding Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with an amber stop codon at amino acid po Svenja Vinke 1800
BBa_K2201263 Coding Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with an amber stop codon at amino acid po Svenja Vinke 1800
BBa_K2201264 Coding Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with an amber stop codon at amino acid po Svenja Vinke 1800
BBa_K2201265 Coding Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with an amber stop codon at amino acid po Svenja Vinke 1800
BBa_K2201266 Coding Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with an amber stop codon at amino acid po Svenja Vinke 1809
BBa_K2201267 Coding Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with an amber stop codon at amino acid po Svenja Vinke 1805
BBa_K2201280 DNA spacer with BseRI recognition site Daniel Bergen 17
BBa_K2201281 DNA spacer with AcuI recognition site Daniel Bergen 20
BBa_K2201282 Coding elastin consensus sequence Daniel Bergen 15
BBa_K2201283 Coding silk consensus sequence Daniel Bergen 18
BBa_K2201300 Coding modeled aminoacyl-tRNA synthetase 2 for the incorporation of CBT-asparagine Denise Kerkhoff 986
BBa_K2201301 Coding modeled aminoacyl-tRNA synthetase 4 for the incorporation of CBT-asparagine Christina Drake 986
BBa_K2201302 Coding modeled aminoacyl-tRNA synthetase 5 for the incorporation of CBT-asparagine Daniel Bergen 986
BBa_K2201331 Generator NM-domain of Sup35 with an amber codon at position 21 under T7-promoter control Laura Schlueter 824
BBa_K2201332 Generator NM-domain of Sup35 with an cysteine at position 21 and 121 and histidine tag. Svenja Vinke 840
BBa_K2201400 Coding Tyrosyl synthetase, library plasmid from Methanococcus jannashii Laura Schlueter 1004
BBa_K2201403 T7 T7RNA Polymerase from E.coli KRX strain. Olga Schmidt 2652
BBa_K2201405 Device T7RNAP+ Olga Schmidt 4008
BBa_K2201406 Translational_Unit Barnase with blanc codons Olga Schmidt 339
BBa_K2201407 Translational_Unit baBL+ Olga Schmidt 1695
BBa_K2201408 Translational_Unit tRNA Olga Schmidt 296
BBa_K2201409 Translational_Unit Kanamycin CDS from pSB1K3 Olga Schmidt 816
BBa_K2201410 Translational_Unit kanRBL+ Olga Schmidt 998
BBa_K2201411 Coding Tyrosyl synthetase library plasmid from M. jannashii with mRFP as optical controle for a selection Laura Schlueter 2003
BBa_K22011414 Coding Kanamycin resistance with two amber stop codon Laura Schlueter 816
BBa_K2201900 Coding Positive selection plasmid for the incorporation of ncAAs Svenja Vinke 1158
BBa_K2201901 Coding Negative selection plasmid for the incorporation of ncAAs Svenja Vinke 681