BBa_K2008007 1 BBa_K2008007 Xylose->comK with AmyE homology 2016-10-08T11:00:00Z 2016-10-18T02:33:48Z This part was amplified from the pAX01-comK plasmid constructed by Zhang and Zhang, 2010, and was previously characterized by the UofC_Calgary 2014 iGEM team. Flanking AmyE regions of homology were derived from the B. subtilis 168 genome. This part is an improvement on the comK gene with the intent to allow the the part to be chromosomally integrated in the B. subtilis genome for increased long-term stability. Internal restriction enzyme sites (one SpeI site located at base pairs 170-175 and two EcoRI sites located at base pairs 557-562 & 572-577 in the comK part submitted by UofC_Calgary 2014) have been removed. In our version of comK, the SpeI site (ACTAGT) was deleted entirely, as the site was contained within a spacer region between the xylose-inducible promoter and RBS. To remove EcoRI sites, the DNA bases of the new comK construct were changed, and codons were optimized for B. subtilis: 553 A>G 556 C>T 568 A>G 571 C>T As the UofC_Calgary 2014 iGEM team discusses in their comK part (BBa_K1444018), comK is the master transcription factor involved in the competency of B. subtilis. ComK further affects the other competence factors (comC, comE, comF, comG and comK) to increase B. subtilis??? ability to be transformed (van Sinderen et al., 1995). As such, when additional comK is transcribed, it increases B. subtilis transformation efficiency. Also like the UofC_Calgary 2014 iGEM team, we included a xylose-inducible promoter upstream of the comK coding sequence. This allows for induction of competency upon the addition of xylose, which is beneficial as the usual starvation method for transformation of B. subtilis is typically time-consuming and costly. Note: This part is a composite part, not a basic part, but it could not be added as composite at the time of entry. false false _2475_ 30207 30207 9 true Two internal EcoRI restriction sites and one internal SpeI restriction site to allow for this part to be BioBrick compatible. Flanking AmyE regions of homology were added to allow for chromosomal integration and increased long-term stability of the comK construct. false Rachelle Varga annotation2489348 1 Stop codon range2489348 1 919 921 annotation2489303 1 5' amyE range2489303 1 1 125 annotation2489346 1 Start codon range2489346 1 343 345 annotation2489349 1 BBa_B0010 range2489349 1 922 1001 annotation2489344 1 PxylA range2489344 1 126 280 annotation2489347 1 comK range2489347 1 346 918 annotation2489345 1 RBS range2489345 1 328 339 annotation2489350 1 BBa_B0012 range2489350 1 1010 1036 annotation2489351 1 3' amyE range2489351 1 1037 1161 BBa_K2008007_sequence 1 tttcatttggttctggcaggaccggcggctgcgagtgctgaaacggcgaacaaatcgaatgagcttacagcaccgtcgatcaaaagcggaaccattcttcatgcatggaattggtcgttcaataccatatctaatattataactaaattttctaaaaaaaacattgaaataaacatttattttgtatatgatgagataaagttagtttattggataaacaaactaactcaattaagatagttgatggataaacttgttcacttaaatcaaagggggaaatgacaaatggtccaagatatctaaaaatcaaagggggaaatgggatccaaaggaggccataatatgagtcagaaaacagacgcacctttagaatcgtatgaagtgaacggcgcaacaattgccgtgctgccagaagaaatagacggcaaaatctgttccaaaattattgaaaaagattgcgtgttttatgtaaacatgaagccgctgcaaattgtcgacagaagctgccgattttttggatcaagctatgcgggaagaaaagcaggaacttatgaagtgacaaaaatttcacacaagccgccgatcatggtggacccttcgaaccaaatctttttattccctacactttcttcgacaagaccccaatgcggctggatttcccatgtgcatgtaaaagagtttaaagcgactgagtttgacgatacggaagtgacgttttccaatgggaaaacgatggagctgccgatctcttataattcgttcgagaaccaggtataccgaacagcgtggctcagaaccaaattccaagacagaatcgaccaccgcgtgccgaaaagacaggaatttatgctgtacccgaaagaagagcggacgaagatgatttatgattttattttgcgtgagctcggggaacggtattagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcccaggcagtacgcctcttttcttttccagacctgagggaggcggaaatggtgtgaggttcccggggaaaagccaaataggcgatcgcgggagtgctttatttgaagatcaggctatcactgcggtc igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z