Team:Saarland/Parts
From 2014.igem.org
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<div id="wikicontent"> | <div id="wikicontent"> | ||
+ | <br> | ||
<h1>Parts:</h1> | <h1>Parts:</h1> | ||
<h3>Hyaluronic acid production</h3> | <h3>Hyaluronic acid production</h3> | ||
- | <h4>Number: | + | <h4>Number: [http://parts.igem.org/Part:BBa_K1469001 ''BBa_K1469001'']</h4> |
- | + | ||
- | + | ||
- | + | ||
- | <b> | + | [[File:PSB1C3 Has2 1.png|thumb|center|450px| <b>Figure 1:</b> vector map of pSB1C3 Has2]] |
- | + | <br> | |
- | + | <b>Description:</b><br> | |
- | + | Hyaluronan synthase 2 gene of<i> Heterocephalus glaber</i> with codon adaption for expression in<i> Bacillus megaterium</i>. The enzyme catalyses the linkage of UDP-D-glucuronic acid and UDP-N-acetyl-D-glucosamine to the linear high molecular mass hyaluronic acid chain and is also involved in its secretion.<br><br> | |
- | + | <html> | |
- | + | <a href="https://2014.igem.org/Team:Saarland/Modeling"> | |
- | + | <img src="https://static.igem.org/mediawiki/2014/d/d2/Links_Pfeil.1.png" width="40px"> | |
- | + | <b> Modelling </b> | |
- | + | </a> | |
- | + | </html> | |
- | + | <br> | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
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- | + | ||
<br> | <br> | ||
+ | <b>Sequence:</b> | ||
+ | <!-- EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469001] --> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGCACTGTGAACGTTTCTTATGTATTTTACGTATTATTGGTACAACATTATTCGGTGTATCTTTATTATTAGGTATTACAGCTGCTTACATTGTAGGTTACCAA | ||
+ | TTCATTCAAACAGATAACTACTACTTCTCTTTCGGTTTATACGGTGCTTTCTTAGCTTCTCACTTAATTATTCAATCTTTATTCGCTTTCTTAGAACACCGTAAA | ||
+ | ATGAAAAAATCTTTAGAAACACCAATTAAATTAAACAAAACAGTAGCTTTATGTATTGCTGCTTACCAAGAAGATCCAGATTACTTACGTAAATGTTTACAATCT | ||
+ | GTAAAACGTTTAACATACCCAGGTATTAAAGTAGTAATGGTAATTGATGGTAACTCTGATGATGATTTATACATGATGGATATTTTCTCTGAAGTAATGGGTCGT | ||
+ | GATAAATCTGCTACATACATTTGGAAAAACAACTTCCACGAAAAAGGTCCAGGTGAAACAGATGAATCTCACAAAGAATCTTCTCAACACGTAACACAATTAGTA | ||
+ | TTATCTTCTAAATCTGTATGTATTATGCAAAAATGGGGTGGTAAACGTGAAGTAATGTACACAGCGTTTCGTGCTCTCGGTCGTTCTGTTGATTACGTACAAGTA | ||
+ | TGTGATTCTGATACAATGTTAGATCCAGCTTCTTCTGTAGAAATGGTAAAAGTATTAGAAGAAGATCCAATGGTAGGTGGTGTAGGTGGTGATGTACAAATTTTA | ||
+ | AACAAATACGATTCTTGGATTTCTTTCTTATCTTCTGTACGTTACTGGATGGCTTTCAACATTGAACGTGCTTGTCAATCTTACTTCGGTTGTGTACAATGTATT | ||
+ | TCTGGTCCATTAGGTATGTACCGTAACTCTTTATTACACGAATTTGTAGAAGATTGGTACTCTCAAGAATTTATGGGTAACCAGTGCAGCTTCGGTGATGATCGT | ||
+ | CACTTAACAAACCGTGTATTATCTTTAGGTTACGCTACAAAATACACAGCTCGTTCTAAATGTTTAACAGAAACACCAATTGAATACTTACGTTGGTTAAACCAA | ||
+ | CAAACACGTTGGTCTAAATCTTACTTCCGTGAATGGTTATACAACGCTATGTGGTTCCACAAACACCACTTATGGATGACATACGAAGCGGTAATAACAGGCTTC | ||
+ | TTCCCATTCTTTTTAATTGCTACAGTAATTCAATTATTCTACCGTGGTAAAATTTGGAACATTTTATTATTCTTATTAACAGTACAATTAGTAGGTTTAATAAAA | ||
+ | TCTTCTTTCGCTTCTTGTTTACGTGGTAACATTGTAATGGTATTCATGTCTTTATACTCTGTATTATACATGTCTTCTTTATTACCAGCTAAAATGTTCGCTATT | ||
+ | GCTACAATTAACAAAGCTGGTTGGGGTACATCTGGTCGTAAAACAATTGTAGTAAACTTCATTGGTTTAATTCCAGTATCTGTATGGTTCACAATTTTATTAGGT | ||
+ | GGTGTAATTTTCACAATTTACAAAGAATCTAAAAAACCATTCTCTGAATCTAAACAAACAGTATTAATTGTAGGTACATTATTATACGCTTGTTACTGGGTAATG | ||
+ | TTATTAACATTATACGTAGTATTAATAAACAAATGTGGTCGTCGTAAAAAAGGTCAACAATACGATATGGTATTAGATGTATAA | ||
+ | <br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
+ | |||
<b>Characterisation status:</b> | <b>Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations | ||
- | *Protein expression successful // | + | *Protein expression successful <html> |
+ | <a href="https://2014.igem.org/Team:Saarland/4_step"> | ||
+ | <img src="https://static.igem.org/mediawiki/2014/d/d2/Links_Pfeil.1.png" width="40px"> | ||
+ | <b> Results </b> | ||
+ | </a> | ||
+ | </html> | ||
*No catalytic activity probably due to misfolding and intracellular accumulation | *No catalytic activity probably due to misfolding and intracellular accumulation | ||
- | <h3>Pathway engineering for | + | <h3>Pathway engineering for optimisation of hyaluronic acid production</h3> |
- | <h4>Number: BBa_K1469002</h4> | + | <h4>Number: [http://parts.igem.org/Part:BBa_K1469002 ''BBa_K1469002'']</h4> |
- | <b>Description:</b> | + | |
- | UDP-glucose 6-dehydrogenase of<i> Bacillus megaterium</i>. The enzyme | + | [[File:PSB1C3 UDP-GlcDH 2.png|thumb|center|450px| <b>Figure 2:</b> vector map of pSB1C3 UDP-GlcDH]] |
- | <b>Sequence:</b> | + | <br> |
- | + | ||
+ | <b>Description:</b><br> | ||
+ | UDP-glucose 6-dehydrogenase of<i> Bacillus megaterium</i>. The enzyme catalyses the NAD dependent oxidation of UDP-glucose to UDP-glucuronic acid, a key precursor molecule for hyaluronic acid production. | ||
+ | <br> | ||
+ | |||
+ | <b>Sequence:</b> <br> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGACTAATATCACAGTAGCGGGTACTGGTTACGTAGGCTTAGTTACAGGTGTTTGTTTAGCTGAAATAGGGCATGAGGTAACATGTGTAGATAA | ||
+ | TATTCAGGATAAAATTGATATTTTAAATGAAGGAATATCTCCTATCTTTGAACCTGGGCTTGATGAATTAATTATTAAAAACTCTGATGCTGGTA | ||
+ | AATTATTTTTCACAACTGATTATCAAAATACTTATGCAAAATCTGATGTTATTTTTATAGGTGTAGGAACACCTGAAAATGAAGATGGGTCTGCT | ||
+ | AATTTAAATTATGTTTATACAGTTGCACAACAAATAGCTAAATTTATTGAGCGAGATTGTCTTATTGTAGTAAAATCTACTGTTCCTATTGGAAC | ||
+ | AAATGAGAAAATAGAAAAATTAATTAAACAACATTTAAAAAATGATGTACAAGTTGAGGTAGCATCCAACCCTGAGTTTTTAGCACAAGGAACAG | ||
+ | CAGTGCGTGACACACTTTATGCATCACGTATTGTTATAGGAGTAGAAAGTGAAAGAGCAAGACAAATATTAACTGATATTTATCAACCTTTTCAT | ||
+ | CAACCTATCTTAACAATGAATCGTCGTAGTGCTGAGATGGTAAAGTATGCTTCAAATGATTTCTTAGCATTAAAAATATCATTTGTTAATGATAT | ||
+ | TGCTAACCTGTGTGAAGTAGTAGGGGCTAACATTGAAGATGTTACAAATGGTATGAGCTATGATCAACGTATTGGAGACAAGTTTTTAAATCCTG | ||
+ | GCATCGGTTACGGTGGTTCATGCTTCCCTAAAGATACTAAAGCGTTACATTGGTTGTCTGAAGAGGAAGGCTATGTATTACGTACCATAAAAGCA | ||
+ | GCTATAGAAGTAAATGAAAAACAAAAGTTTAAACTGATTAAAAAAGCAAGACAAGACTTTGCAACTTTCAAGGGTTTAAAAGTTGCTGTTTTAGG | ||
+ | TCTTACATTCAAGCCTGGTACCGATGACTTACGGGAGGCACCATCAATTCCCAATATTCGTTTATTATTAAATGAAGGTGCTGAAGTTCATGCAT | ||
+ | ATGATCCTGTAGGTGTAAACAATTTTAAGAAAATATATCCAACACAAGTGTATTATGAAGAAACAATTGAAAAAGCATTAAAAAATGCTGACATT | ||
+ | TGTTTTATATTTACAGAGTGGGAAGAAATAAAAAATATGAATTTAAACTTGTTTAACAAACAAATGAAATCTCCTTTTGTCTATGATGGACGTAA | ||
+ | TTGTTACAATCTCGATGAAGTTAAAAGAAGTGGTATCAATTATCAATCTATTGGTCGTCCTAATATAAATGGATACAAAGCTAAAATTAAAGCTA<br> | ||
+ | ATGTCTAA<br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
<b>Characterisation status:</b> | <b>Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations | ||
- | *Protein expression successful // | + | *Protein expression successful <html> |
+ | <a href="https://2014.igem.org/Team:Saarland/4_step#WesternBlot"> | ||
+ | <img src="https://static.igem.org/mediawiki/2014/d/d2/Links_Pfeil.1.png" width="40px"> | ||
+ | <b> SDS PAGE</b> | ||
+ | </a> | ||
+ | </html> | ||
*characterisation of catalytic activity not possible because no HA production for comparison of HA yields after co expression with hyaluronan synthase 2. | *characterisation of catalytic activity not possible because no HA production for comparison of HA yields after co expression with hyaluronan synthase 2. | ||
- | |||
- | < | + | <h4>Number: [http://parts.igem.org/Part:BBa_K1469003 ''BBa_K1469003'']</h4> |
- | + | ||
- | + | ||
- | + | ||
+ | [[File:PSB1C3 Glk 3.png|thumb|center|450px| <b>Figure 3:</b> vector map of pSB1C3 Glk]] | ||
+ | <br> | ||
+ | |||
+ | <b>Description:</b><br> | ||
+ | Glucokinase of <i>Bacillus megaterium</i>. The enzyme catalyses the phosphorylation reaction of glucose to glucose 6-phospahate.<br> | ||
+ | |||
+ | <b>Sequence: </b> <br> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGGATGACAAATGGTTAGTTGGAGTTGATTTAGGCGGTACAACAATAAAAATGGCCTTTATTAATCATTATGGAGAAATCATTCACAAGTGGA | ||
+ | AATTAATACGGATGTGAGCGAGCAAGGCCGTAAAATTCCAACGGATATTGCAAAAGCAATTGATAAAAAGTTAAACGATCTTGGAGAAGTAAAA | ||
+ | TCAAGGTTAGTAGGAATTGGCATTGGTGCACCGGGGCCTGTCAACTTTGCAAACGGTTCGATTGAAGTAGCTGTCAATTTAGGTTGGGAAAAAT | ||
+ | TCCCTATAAAAGATATCTTGGAAGTAGAAACTTCTCTTCCTGTTGTAGTAGACAATGATGCAAACATTGCAGCGATTGGAGAAATGTGGAAGGG | ||
+ | TGCTGGAGACGGAGCAAAAGATTTACTTTGCGTTACGCTTGGCACAGGCGTTGGCGGTGGCGTCATTGCAAACGGTGAAATTGTACAAGGCGTA | ||
+ | AATGGAGCCGCTGGTGAGATCGGGCACATTACTTCTATTCCTGAAGGCGGGGCACCGTGTAACTGCGGTAAAACCGGCTGTTTAGAAACCATTG | ||
+ | CTTCAGCAACTGGAATTGTACGTTTAACAATGGAAGAATTAACGGAAACGGACAAACCAAGTGAGCTTCGCACAGTGTTAGAACAAAATGGACA | ||
+ | AGTTACATCTAAAGATGTATTTGATGCAGCTCGTTCAAAAGACGGGTTAGCTATGCATGTTGTAGATAAAGTTGCTTTTCATTTAGGTCTAGCA | ||
+ | CTAGCAAACTCTGCTAATGCATTAAACCCTGAGAAGATCGTTCTAGGCGGCGGTGTGTCTCGTGCAGGCGAGGTATTACTTGCACCGGTAAGAG | ||
+ | ATTATTTCAAACGTTTTGCATTTCCTCGCGTAGCGCAAGGTGCTGAACTAGCAATCGCTACTTTAGGAAACGATGCGGGAATTATTGGAGGAGC | ||
+ | TTGGTTAGTTAAATCTTATTTTGAATAA<br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
<b>Characterisation status:</b> | <b>Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations | ||
Line 65: | Line 133: | ||
- | <h4>Number: | + | <h4>Number: [http://parts.igem.org/Part:BBa_K1469004 ''BBa_K1469004'']</h4> |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
+ | [[File:PSB1C3 PgmB 4.png|thumb|center|450px| <b>Figure 4:</b> vector map of pSB1C3 pgmB]] | ||
+ | <br> | ||
+ | |||
+ | <b>Description:</b> <br> | ||
+ | Beta-phosphoglucomutase of <i>Bacillus megaterium</i>. The enzyme catalyses the conversion of glucose 6-phospahate to glucose 1-phosphate.<br> | ||
+ | |||
+ | <b>Sequence:</b><br> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGAAAAAAGAAGAGCTACAAGCGATTATTTTTGATTTAGACGGTGTTATTGCAGATACGGTACATTTATATTATAAAGCAAATAAAAAAGT | ||
+ | AGCAGATCAACTTAACGTTTCATTTTCTGAAGAACTCAATCAGCAGCTGCAGGGGATCAGCCGAGTGAAAACTGTAGAGCTCATTGCTGCCC | ||
+ | AAGGCAATGTACATCTGACAATGGAAAAAAAAGAACAGCTTGCTGATGAAAAGAATAAGCATTATCAAGCATTAATAAAGGAAATGACAAAG | ||
+ | GATCATCTACTTCCAGGAATACAATTGTTGTTAGAGGATTGTAAAAAACAAGGTATTAAAATGGCGATTGCTTCTTCTAGTTCAAATGCAAA | ||
+ | TACTGTAGTGGATGCGTTAGGAATACGTCCTTTTTTTGATTGCATTGTGGATGTGAGAAAAATAAAGAAGGGAAAACCAGACCCAGAAATAT | ||
+ | TTTTGACAGCTGCCCATCAATTAAAAGTAACACCCTATGCATGTGTGGCTATAGAAGATGGAGAGGCGGGTGTAAAAGCTATTAATCAAACA | ||
+ | GATATGGTTTCTATCGGCGTTGGTAAGCATCTGAAGGAAGAGGATATGAATTTTCACGTATATTCAACAGCTGATTTGCATTTAAAGACGAT | ||
+ | CAAAGAGGTTTTCGAAAAAACAAGAAAAGGAACAGATTATAACAGCGTGTATACAAAAAATTAA<br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
<b>Characterisation status:</b> | <b>Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations | ||
Line 77: | Line 161: | ||
*Use only after amplification with appropriate primers | *Use only after amplification with appropriate primers | ||
- | <h4>Number: BBa_K1469005 </h4> | + | |
- | <b>Description:</b> | + | <h4>Number: [http://parts.igem.org/Part:BBa_K1469005''BBa_K1469005''] </h4> |
- | UTP--glucose-1-phosphate uridylyltransferase of <i>Bacillus megaterium</i>. The enzyme | + | |
- | <b>Sequence:</b> | + | [[File:PSB1C3 GtaB 5.png|thumb|center|450px| <b>Figure 5:</b> vector map of pSB1C3 gtaB]] |
- | + | <br> | |
+ | |||
+ | <b>Description:</b><br> | ||
+ | UTP--glucose-1-phosphate uridylyltransferase of <i>Bacillus megaterium</i>. The enzyme catalyses the conversion of glucose 1-phospahate to UDP-glucose 1-phosphate.<br> | ||
+ | |||
+ | <b>Sequence:</b> <br> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGACGATAAAAAAGGCAGTTATTCCAGTAGGCGGTTTAGGAACTCGATTTTTGCCAGCGACAAAAGCGCAGCCCAAAGAAATGCTGCCAAT | ||
+ | TGTCGATAAGCCTGCTGTACAATATATTGTAGAAGAAGCTGTGAAATCAGGTATTGAAAGTATTATTTTTATTACAGGACGAAACAAAAAAT | ||
+ | CAATTGAAGACCATTTTGATAAATCAACTGAGTTAGAGCAAATGCTTGAAGAAAAACACAAGTTCCAAATGCTAAAAGAAGTACAAACGATC | ||
+ | TCATCAATGGCCAGCATTCACTATATTCGTCAAAAAGAGCCGTTAGGACTTGGTCATGCTATTCTTTGTGCCGAACAGTTTATTGGAGATGA | ||
+ | GCCGTTTGCCGTTCTTCTTGGAGACGATATTATGACTTCTGAAGAACCTGCGCTCAAACAGATAATCCAAGCGTATGAAACGACTAATCAAT | ||
+ | CAGTTATCGGAGTTCAAAAAGTGGATGTTGAAGAGGTATCAAAATACGGAATCATTCAGCCGAAAAATAAATCTGGCAGCCTTCATGAAGTG | ||
+ | AATGACTTGATTGAAAAACCATCGATAGAACAAGCTCCGTCAACTATTGCCGTTATGGGCCGCTATATTTTAAATCCTTCTATTTTTTCTTA | ||
+ | TCTAAAAACGATTGAAAGAGGGGTAGGAAATGAATTACAATTAACAGATGCACTGGGTGTGGTATGTGAAAAAGAGCGATTGTTTGCTCTTG | ||
+ | AACTGGAAGGACAGCGCTTTGATATTGGAGATAAGGTAGGCTATATGAAAGCGATGGTAGAAGTTGCGTTAAAGCGCAGAGATTTACGTCAA | ||
+ | ACGTTTTTATCGTATTTGGAGGGAATTGTAGAAAAAGAACGAGCAAACAACTTCAGCTAA<br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
<b>Characterisation status:</b> | <b>Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations | ||
Line 88: | Line 192: | ||
*Use only after amplification with appropriate primers | *Use only after amplification with appropriate primers | ||
- | |||
- | |||
- | |||
- | < | + | <h4>Number: [http://parts.igem.org/Part:BBa_K1469006 ''BBa_K1469006'']</h4> |
- | + | ||
- | + | [[File:PSB1C3 Pgi 6.png|thumb|center|450px| <b>Figure 6:</b> vector map of pSB1C3 pgi]] | |
+ | <br> | ||
+ | |||
+ | <b>Description:</b><br> | ||
+ | Glucose-6-phosphate isomerase of <i>Bacillus megaterium</i>. The enzyme catalyses the conversion of glucose 6-phospahate to fructose 6-phosphate.<br> | ||
+ | |||
+ | <b>Sequence:</b> <br> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGACACATGTTCGTTTTGATTATTCAAACGCGTTATCATTTTTTGGTGAACATGAACTTACATACTTACGTGATGCCGTGAAAGTAGCTC | ||
+ | ACCATTCAATTCATGAGAAAACAGGTGCTGGAAACGACTTTTTAGGATGGGTCGACCTTCCTACGGAGTATGACAAAGAAGAATTTGCACG | ||
+ | CATTCAAAAAAGTGCAGAAAAAATTAAATCTGATTCAGATGTATTAATTGTTATCGGTATTGGTGGATCTTACTTAGGTGCTCGTGCTGCT | ||
+ | ATTGAAATGCTTCAGCATTCATTCTATAACACATTATCAAAAGAACAGCGTAAAACTCCGCAAGTTATTTTTGCTGGTAACAACATCAGCT | ||
+ | CAACATACATGAAAGACTTAATGGATGTTTTACAAGATAAAGATTTCTCTATTAATGTTATTTCAAAATCTGGTACAACAACAGAACCGGC | ||
+ | GATTGCTTTCCGTATCTTCCGTAAATTATTAGAAGAGAAATACGGCAAAGAAGAAGCACGCAAGCGTATTTATGCAACAACAGATAAAGCG | ||
+ | CGCGGTGCATTAAAAACATTAGCGGACGAAGAAGGCTATGAAACATTTGTTATTCCAGATGATGTAGGCGGCCGTTATTCAGTATTAACAG | ||
+ | CGGTTGGCTTATTACCAATTGCCGCTGCGGGTCTTGATATCGAACAAATGATGAAAGGGGCAGCTGATGCTCAAGAAGCATTCTCATCATC | ||
+ | AGAGCTAGAAGAGAACCCAGCTTATCAATATGCTGTAGCTCGTAATGTGCTTTATAATAAAGGAAAAACAATTGAAATGCTGATTAACTAT | ||
+ | GAGCCAGCTCTTCAATACTTTGCTGAGTGGTGGAAACAGTTATTTGGCGAAAGTGAAGGAAAAGACCAAAAAGGTATTTACCCTTCATCTG | ||
+ | CTAACTTCTCAACGGATCTTCATTCTTTAGGTCAATATGTACAAGAAGGTCGCCGTGATATTTTTGAAACAATTGTACAAGTTCAAGATTC | ||
+ | TCGTCATGAGCTAACAATTGAAGAAGATGCAGCGGACTTAGATGGATTAAATTACTTAGCTGGTGAAACAGTTGACTTTGTAAATAAAAAA | ||
+ | GCGTTCCAAGGTACAATGCTTGCTCACACAGATGGAGGCGTTCCAAACTTAGTTGTGAACATTCCTGCGCTAGATGAGTATACGTTCGGAT | ||
+ | ATGTTGTATATTTCTTCGAAAAAGCATGTGCAATCAGCGGCTATTTATTAGGCGTAAACCCATTCGATCAGCCGGGAGTAGAAGCATACAA | ||
+ | AGTAAATATGTTTGCTCTTCTTGGAAAACCTGGTTTTGAAGAGAAAAAAGCAGAATTAGAAAAGCGTTTGAAATAA<br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
+ | <b>Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations | ||
*Part is not RFC10 conform | *Part is not RFC10 conform | ||
Line 101: | Line 229: | ||
- | <h4>Number: BBa_K1469007</h4> | + | <h4>Number: [http://parts.igem.org/Part:BBa_K1469007 ''BBa_K1469007'']</h4> |
- | <b>Description:</b> | + | |
- | Glucosamine--fructose-6-phosphate aminotransferase (isomerising) of <i>Bacillus megaterium</i>. The enzyme | + | [[File:PSB1C3 GlmS 7.png|thumb|center|450px| <b>Figure 7:</b> vector map of pSB1C3 glmS]] |
- | <b>Sequence:</b> | + | <br> |
- | + | ||
- | + | <b>Description:</b><br> | |
+ | Glucosamine--fructose-6-phosphate aminotransferase (isomerising) of <i>Bacillus megaterium</i>. The enzyme catalyses the conversion of fructose 6-phosphate to glucosamine 6-phosphate.<br> | ||
+ | |||
+ | <b>Sequence:</b> <br> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGTGCGGAATTGTAGGATATATTGGAACAGAAGATTCGAAAGAAATTTTATTACGTGGTCTTGAAAAGTTAGAATATCGTGGCTATGACT | ||
+ | CAGCGGGTATTGCAGTTGTTAACAATGAAGGCGTACACGTATTCAAAGAAAAAGGCCGTATTGCAGAATTGCGTCAGGCAGTAGATAACAG | ||
+ | TGTTGTAGCACCAGCAGGTATCGGTCATACGCGTTGGGCAACTCATGGTGTACCAAGCCAAGAAAATGCTCACCCGCATCAAAGTACATCT | ||
+ | GGACGCTTCACACTTGTGCATAACGGTGTAATTGAAAACTATGCTATTTTGAAACGTGAATATTTACAAGATGTAACGTTCAAAAGCGAAA | ||
+ | CGGATACAGAAGTAATCGTTCAGTTAATTCAAAAAATCGCAGCAGAAGGCTTAGATGTAGAAGAAGCGTTCCGTAAGACAGTAAGCTTGCT | ||
+ | TCATGGTTCTTATGCACTAGCTCTTGTTGATAACAAAGATAAAAATACAATCTACGTAGCTAAAAACAAAAGCCCGCTTCTTGTAGGTGTT | ||
+ | GGTGAAGGATTTAACGTTGTAGCGAGTGATGCTATGGCAATGCTTCAAGTAACAGATCAGTACGTAGAATTAATGGATAAAGAAACAGTAA | ||
+ | TTGTTACAAAAGAAGGCGTAACAATTAAAACGCTTGATGGTGAAGTGGTAGAAAGAGCACCTTATACAGCAGAGCTTGATGCAAGTGATAT | ||
+ | TGAAAAAGGTACGTACCCTCACTATATGTTAAAAGAAATTGACGAGCAGCCTTTAGTTGTTCGTAAAATCATTCAAGAGTATCAAAATGAG | ||
+ | AACAATGAACTGCACATTGATGCAGACATTTTAGGAGCAATGGATGAAGCAGACCGCATCTATATCGTAGCATGTGGAACAAGCTATCATG | ||
+ | CAGGCCTAGTAGGTAAACAATTCATTGAAACGTGGGCGAAGGTTCCAGTAGAAGTGCACGTAGCAAGTGAATTTTCTTATAACATGCCGCT | ||
+ | TTTATCTGAAAAGCCATTGTTTATCTTTATTTCTCAAAGTGGAGAAACAGCTGACAGCCGTGCAGTATTAGTTCAAATTAAAGAACTAGGT | ||
+ | TACAAAGCGTTAACAATCACAAACGTTCCGGGATCAACATTATCTCGTGAATCTGATTATACATTATTACTTCATGCAGGTCCGGAAATTG | ||
+ | CTGTAGCATCAACAAAAGCTTATACAGCTCAATTAGCAGTGCTATCAATTCTAGCGGCTGTAACAGCAAAAACACGCGGCATTGAATTAGA | ||
+ | ATTTGATTTAGTTCAAGAGTTAGCAATCGTAGCAAACACAATGGAAGTACTTTGCGACGACAAAGAAGAAATGGAAAGCATTGCTCTTCAA | ||
+ | TTCTTAGCAACAACTCGCAACGCATTCTTTATTGGACGTTCTGTCGATTATTATGTAGGTTTAGAAGGTGCGTTAAAACTAAAAGAAATCT | ||
+ | CTTACATCCAAGCAGAAGGATTTGCTGGAGGAGAGCTTAAGCACGGAACAATCGCTTTAATTGAAAACAATACGCCAATTATTGCATTAGC | ||
+ | AACACAAGAGCATGTAAACTTAAGCATCCGTGGTAACGTAAAAGAAGTAGTAGCACGTGGAGCTAATCCTTGTATCATTTCAATGAAAGGA | ||
+ | TTAGAAACAGAAGAAGATCGCTACGTGATTCCGAAAGTTCACGAAACACTTTCACCACTAGCAGCGGTTATTCCTTTACAATTAATCTCTT | ||
+ | ACTACGCTGCATTGCACCGCGATTGCGATGTAGATAAACCACGTAACTTAGCTAAATCAGTTACTGTAGAATAA<br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
<b> Characterisation status:</b> | <b> Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations | ||
Line 113: | Line 269: | ||
*Use only after amplification with appropriate primers | *Use only after amplification with appropriate primers | ||
- | |||
- | |||
- | |||
- | |||
- | |||
+ | <h4>Number: [http://parts.igem.org/Part:BBa_K1469008 ''BBa_K1469008'']</h4> | ||
+ | |||
+ | [[File:PSB1C3 GlmM 8.png|thumb|center|450px| <b>Figure 8:</b> vector map of pSB1C3 glmM]] | ||
+ | <br> | ||
+ | |||
+ | <b>Description:</b><br> | ||
+ | Phosphoglucosamine mutase of <i>Bacillus megaterium</i>. The enzyme catalyses the conversion of glucosamine 6-phosphate to glucosamine 1-phosphate.<br> | ||
+ | |||
+ | <b>Sequence:</b><br> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGGGTAAGTATTTTGGAACAGACGGAGTACGAGGAGTTGCCAATAGTGAATTAACTCCTGAGCTTGCTTTTAAAATTGGACGCTTAGGTG | ||
+ | GTTACGTTCTAACAAAAGATTCAGAGCGACCAAAAGTATTAATCGGACGCGATACACGCGTTTCTGGTCATATGTTAGAAGGAGCGCTTGT | ||
+ | TGCAGGATTGCTTTCAATTGGAGCAGAAGTTATGCGACTAGGTGTTATTTCAACTCCAGGGGTAGCGTATTTAACAAAAGCACTTGGCGCA | ||
+ | CAAGCAGGAGTCATGATTTCTGCTTCGCATAACCCAGTTCAAGATAACGGAATTAAATTCTTTGGTCCAGATGGATTTAAACTTTCAGATG | ||
+ | CCCAAGAGAATGAAATTGAAGCGTTAATGGATAGTGACACAGATCAATTACCAAGACCTGTTGGGGGAGACTTAGGTCAGGTAAATGACTA | ||
+ | TTTCGAAGGTGGACAAAAATATCTTCAATACTTAAAACAAACGGTTGATGAAGACTTCTCAGGTATTCATGTGGCATTAGACTGTGCGCAC | ||
+ | GGGGCAACGTCATCGTTAGCAGCACATTTATTTGCAGATCTCGATGCAGATATCTCTACAATGGGGGCATCTCCAAACGGATTGAACATCA | ||
+ | ATGACGGTGTAGGATCTACACATCCGGAAGCATTAAGTGCCTTCTTAAAAGAAAAAGGAGCGGATGTTGGATTAGCATTTGATGGAGACGG | ||
+ | AGATCGTTTAATCGCAATCGACGAAAAAGGCGAGATTGTAGACGGTGATCAAATTATGTTCATTTGTGCAAAATACTTATATGCACAAGGT | ||
+ | CGTTTAAAGAAAAACACGCTTGTTTCTACCGTTATGAGTAACTTAGGCTTCTATAAAGCACTAGAAGCTAGCGGGATTGAAAGTGCACAAA | ||
+ | CTGGTGTAGGAGACCGTTATGTAGTAGAAGAAATGAAAGCTAATCAATATAACCTAGGTGGAGAACAGTCAGGTCATATTATTTTCTTAGA | ||
+ | TTACAACACAACAGGTGACGGTATGCTTTCAGCTATTCAGCTTGTAAACATTATGCAAGCAACGAAAAAGCCTTTATCAGAATTAGCAGCT | ||
+ | GAAATGAAGAAATATCCTCAGCTTCTTGTAAATGTAAAAGTAACAGATAAGCATCATGTTACAGATAACGAGAAGGTAAAAGTGGTTATTG | ||
+ | AAGAAGTGGAAAAAGAAATGAACGGTAATGGCCGAGTACTTGTACGCCCTTCAGGAACAGAGCCGTTAGTACGTGTGATGGTTGAAGCACA | ||
+ | AACACAAGAAGAATGTGAAACTTATGTAACGCGTATCGTAGAAGTAGTAAAAGAAGAAATGGGCTTAGAGTAA<br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
<b>Characterisation status:</b> | <b>Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations | ||
Line 125: | Line 305: | ||
*Use only after amplification with appropriate primers | *Use only after amplification with appropriate primers | ||
- | <h4>Number: BBa_K1469009</h4> | + | |
- | <b>Description:</b> | + | <h4>Number: [http://parts.igem.org/Part:BBa_K1469009 ''BBa_K1469009'']</h4> |
- | UDP-N-acetylglucosamine pyrophosphorylase of <i>Bacillus megaterium</i>. The enzyme is bifunctional and | + | |
- | <b>Sequence:</b> | + | [[File:PSB1C3 GcaD neu.png|thumb|center|450px| <b>Figure 9:</b> vector map of pSB1C3 gcaD]] |
- | + | <br> | |
+ | |||
+ | <b>Description:</b><br> | ||
+ | UDP-N-acetylglucosamine pyrophosphorylase of <i>Bacillus megaterium</i>. The enzyme is bifunctional and catalyses the conversion of glucosamine 1-phosphate to N-acetyl glucosamine 1-phosphate as well as the conversion of N-acetyl glucosamine 1-phosphate to UDP-N-acetyl glucosamine, a key precursor molecule for hyaluronic acid production.<br> | ||
+ | |||
+ | <b>Sequence:</b> <br> | ||
+ | <html> | ||
+ | <span style="font-family:Courier New"> | ||
+ | ATGTCAAAAAGATATGCAGTCATATTGGCAGCTGGTCAGGGTACACGTATGAAGTCATCTTTATATAAAGTGTTACACCCTGTGTGTGGCAA | ||
+ | ACCGATGGTGCAACACGTAATTGATCAGTTATCACGTTTGGATTTAACAAGTTTAATTACAGTTGTAGGTCACGGTGCCGAAAAAGTGAAAT | ||
+ | CACATGTTGGAGATAAAAGCTTATTTGCTTTACAAGAAGAGCAGTTAGGAACAGCTCATGCAGTAATGCAAGCAGAATCTTTACTTGCACAT | ||
+ | GAAAAAGGTACGACAATTGTTGTATGTGGAGATACTCCGCTTATTACAGCTGAAACAATTGAATCTTTATTAAAGCATCATGAAGCAACAAA | ||
+ | TGCAATGGCAACTATTTTAACAGCTCATGCAGAAGATCCAACAGGATATGGACGCATCATTCGAAATGAACAAGGGTCTGTTGAAAAGATTG | ||
+ | TGGAGCATAAAGATGCCACAGAACAAGAGCGTCTTGTACAGGAAATTAATACTGGTACGTATTGTTTTGATAATGAGAAATTATTTGAGACA | ||
+ | CTTTCAAAAGTTTCAAATGATAATGTACAGGGAGAGTATTATTTACCTGATGTTATCGAAATTCTAAAAAATGAAGGTCAAATTATTTCTGC | ||
+ | ATATCAAACATCCGATTTTGCTGAAACATTGGGCGTTAATGATCGATTTGCTCTGTCTCAAGCAGAAGAAACAATGAAAAAGCGCATCAACA | ||
+ | AGCAGCATATGTTAAATGGAGTGACGATTGTGGATCCTTCTAACACGTACATTTCAGCAGATGCTGTCATTGGCAGAGATACGGTTATTTAT | ||
+ | CCTGGTACGGTTATTCAAGGAACAGTTGTAATTGGAGAGAATTGTGAAGTAGGACCTAACAGTGAAATTAAAGACTGCAAAATAGGTAATAA | ||
+ | TACTTCTATTCGCCATTCTGTTGCTCACGATAGTGAAATTGGTCACGAAGTCACAATTGGCCCATTTGCACATATTCGTCCTCAATCATTAA | ||
+ | TTGGAGATGAAGTGCGCGTAGGAAACTTTGTAGAAATTAAAAAAGCATCTTTTGGCAAAGGTAGCAAAGCTTCTCATTTAAGTTATATTGGA | ||
+ | GACGCAGAGGTCGGAAAAGGTGTAAACTTAGGGTGCGGATCTATTACAGTTAACTACGATGGGAAAAATAAATTTTTAACAAAGATTGAAGA | ||
+ | TGGGGCATTCGTAGGATGTAACTCAAACTTAATTGCACCTGTAACAATTGGTGAAGGCGCCTATGTTGCAGCTGGTTCAACCGTAACGGATG | ||
+ | ATGTGCCAGGAAAAGCATTATCAATTGCTCGTGCGAAACAAGTAAATAAAGAAAACTATGCTGAAAAGCTTGATATTAATAAAAAATCCTAA<br> | ||
+ | </span> | ||
+ | </html> | ||
+ | <br> | ||
<b>Characterisation status:</b> | <b>Characterisation status:</b> | ||
*Part is user sequenced: no illegal restriction sites, no mutations | *Part is user sequenced: no illegal restriction sites, no mutations |
Latest revision as of 12:14, 17 October 2014
Parts:
Hyaluronic acid production
Number: [http://parts.igem.org/Part:BBa_K1469001 BBa_K1469001]
Description:
Hyaluronan synthase 2 gene of Heterocephalus glaber with codon adaption for expression in Bacillus megaterium. The enzyme catalyses the linkage of UDP-D-glucuronic acid and UDP-N-acetyl-D-glucosamine to the linear high molecular mass hyaluronic acid chain and is also involved in its secretion.
Modelling
Sequence:
ATGCACTGTGAACGTTTCTTATGTATTTTACGTATTATTGGTACAACATTATTCGGTGTATCTTTATTATTAGGTATTACAGCTGCTTACATTGTAGGTTACCAA
TTCATTCAAACAGATAACTACTACTTCTCTTTCGGTTTATACGGTGCTTTCTTAGCTTCTCACTTAATTATTCAATCTTTATTCGCTTTCTTAGAACACCGTAAA
ATGAAAAAATCTTTAGAAACACCAATTAAATTAAACAAAACAGTAGCTTTATGTATTGCTGCTTACCAAGAAGATCCAGATTACTTACGTAAATGTTTACAATCT
GTAAAACGTTTAACATACCCAGGTATTAAAGTAGTAATGGTAATTGATGGTAACTCTGATGATGATTTATACATGATGGATATTTTCTCTGAAGTAATGGGTCGT
GATAAATCTGCTACATACATTTGGAAAAACAACTTCCACGAAAAAGGTCCAGGTGAAACAGATGAATCTCACAAAGAATCTTCTCAACACGTAACACAATTAGTA
TTATCTTCTAAATCTGTATGTATTATGCAAAAATGGGGTGGTAAACGTGAAGTAATGTACACAGCGTTTCGTGCTCTCGGTCGTTCTGTTGATTACGTACAAGTA
TGTGATTCTGATACAATGTTAGATCCAGCTTCTTCTGTAGAAATGGTAAAAGTATTAGAAGAAGATCCAATGGTAGGTGGTGTAGGTGGTGATGTACAAATTTTA
AACAAATACGATTCTTGGATTTCTTTCTTATCTTCTGTACGTTACTGGATGGCTTTCAACATTGAACGTGCTTGTCAATCTTACTTCGGTTGTGTACAATGTATT
TCTGGTCCATTAGGTATGTACCGTAACTCTTTATTACACGAATTTGTAGAAGATTGGTACTCTCAAGAATTTATGGGTAACCAGTGCAGCTTCGGTGATGATCGT
CACTTAACAAACCGTGTATTATCTTTAGGTTACGCTACAAAATACACAGCTCGTTCTAAATGTTTAACAGAAACACCAATTGAATACTTACGTTGGTTAAACCAA
CAAACACGTTGGTCTAAATCTTACTTCCGTGAATGGTTATACAACGCTATGTGGTTCCACAAACACCACTTATGGATGACATACGAAGCGGTAATAACAGGCTTC
TTCCCATTCTTTTTAATTGCTACAGTAATTCAATTATTCTACCGTGGTAAAATTTGGAACATTTTATTATTCTTATTAACAGTACAATTAGTAGGTTTAATAAAA
TCTTCTTTCGCTTCTTGTTTACGTGGTAACATTGTAATGGTATTCATGTCTTTATACTCTGTATTATACATGTCTTCTTTATTACCAGCTAAAATGTTCGCTATT
GCTACAATTAACAAAGCTGGTTGGGGTACATCTGGTCGTAAAACAATTGTAGTAAACTTCATTGGTTTAATTCCAGTATCTGTATGGTTCACAATTTTATTAGGT
GGTGTAATTTTCACAATTTACAAAGAATCTAAAAAACCATTCTCTGAATCTAAACAAACAGTATTAATTGTAGGTACATTATTATACGCTTGTTACTGGGTAATG
TTATTAACATTATACGTAGTATTAATAAACAAATGTGGTCGTCGTAAAAAAGGTCAACAATACGATATGGTATTAGATGTATAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Protein expression successful
Results
- No catalytic activity probably due to misfolding and intracellular accumulation
Pathway engineering for optimisation of hyaluronic acid production
Number: [http://parts.igem.org/Part:BBa_K1469002 BBa_K1469002]
Description:
UDP-glucose 6-dehydrogenase of Bacillus megaterium. The enzyme catalyses the NAD dependent oxidation of UDP-glucose to UDP-glucuronic acid, a key precursor molecule for hyaluronic acid production.
Sequence:
ATGACTAATATCACAGTAGCGGGTACTGGTTACGTAGGCTTAGTTACAGGTGTTTGTTTAGCTGAAATAGGGCATGAGGTAACATGTGTAGATAA
TATTCAGGATAAAATTGATATTTTAAATGAAGGAATATCTCCTATCTTTGAACCTGGGCTTGATGAATTAATTATTAAAAACTCTGATGCTGGTA
AATTATTTTTCACAACTGATTATCAAAATACTTATGCAAAATCTGATGTTATTTTTATAGGTGTAGGAACACCTGAAAATGAAGATGGGTCTGCT
AATTTAAATTATGTTTATACAGTTGCACAACAAATAGCTAAATTTATTGAGCGAGATTGTCTTATTGTAGTAAAATCTACTGTTCCTATTGGAAC
AAATGAGAAAATAGAAAAATTAATTAAACAACATTTAAAAAATGATGTACAAGTTGAGGTAGCATCCAACCCTGAGTTTTTAGCACAAGGAACAG
CAGTGCGTGACACACTTTATGCATCACGTATTGTTATAGGAGTAGAAAGTGAAAGAGCAAGACAAATATTAACTGATATTTATCAACCTTTTCAT
CAACCTATCTTAACAATGAATCGTCGTAGTGCTGAGATGGTAAAGTATGCTTCAAATGATTTCTTAGCATTAAAAATATCATTTGTTAATGATAT
TGCTAACCTGTGTGAAGTAGTAGGGGCTAACATTGAAGATGTTACAAATGGTATGAGCTATGATCAACGTATTGGAGACAAGTTTTTAAATCCTG
GCATCGGTTACGGTGGTTCATGCTTCCCTAAAGATACTAAAGCGTTACATTGGTTGTCTGAAGAGGAAGGCTATGTATTACGTACCATAAAAGCA
GCTATAGAAGTAAATGAAAAACAAAAGTTTAAACTGATTAAAAAAGCAAGACAAGACTTTGCAACTTTCAAGGGTTTAAAAGTTGCTGTTTTAGG
TCTTACATTCAAGCCTGGTACCGATGACTTACGGGAGGCACCATCAATTCCCAATATTCGTTTATTATTAAATGAAGGTGCTGAAGTTCATGCAT
ATGATCCTGTAGGTGTAAACAATTTTAAGAAAATATATCCAACACAAGTGTATTATGAAGAAACAATTGAAAAAGCATTAAAAAATGCTGACATT
TGTTTTATATTTACAGAGTGGGAAGAAATAAAAAATATGAATTTAAACTTGTTTAACAAACAAATGAAATCTCCTTTTGTCTATGATGGACGTAA
TTGTTACAATCTCGATGAAGTTAAAAGAAGTGGTATCAATTATCAATCTATTGGTCGTCCTAATATAAATGGATACAAAGCTAAAATTAAAGCTA
ATGTCTAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Protein expression successful
SDS PAGE
- characterisation of catalytic activity not possible because no HA production for comparison of HA yields after co expression with hyaluronan synthase 2.
Number: [http://parts.igem.org/Part:BBa_K1469003 BBa_K1469003]
Description:
Glucokinase of Bacillus megaterium. The enzyme catalyses the phosphorylation reaction of glucose to glucose 6-phospahate.
Sequence:
ATGGATGACAAATGGTTAGTTGGAGTTGATTTAGGCGGTACAACAATAAAAATGGCCTTTATTAATCATTATGGAGAAATCATTCACAAGTGGA
AATTAATACGGATGTGAGCGAGCAAGGCCGTAAAATTCCAACGGATATTGCAAAAGCAATTGATAAAAAGTTAAACGATCTTGGAGAAGTAAAA
TCAAGGTTAGTAGGAATTGGCATTGGTGCACCGGGGCCTGTCAACTTTGCAAACGGTTCGATTGAAGTAGCTGTCAATTTAGGTTGGGAAAAAT
TCCCTATAAAAGATATCTTGGAAGTAGAAACTTCTCTTCCTGTTGTAGTAGACAATGATGCAAACATTGCAGCGATTGGAGAAATGTGGAAGGG
TGCTGGAGACGGAGCAAAAGATTTACTTTGCGTTACGCTTGGCACAGGCGTTGGCGGTGGCGTCATTGCAAACGGTGAAATTGTACAAGGCGTA
AATGGAGCCGCTGGTGAGATCGGGCACATTACTTCTATTCCTGAAGGCGGGGCACCGTGTAACTGCGGTAAAACCGGCTGTTTAGAAACCATTG
CTTCAGCAACTGGAATTGTACGTTTAACAATGGAAGAATTAACGGAAACGGACAAACCAAGTGAGCTTCGCACAGTGTTAGAACAAAATGGACA
AGTTACATCTAAAGATGTATTTGATGCAGCTCGTTCAAAAGACGGGTTAGCTATGCATGTTGTAGATAAAGTTGCTTTTCATTTAGGTCTAGCA
CTAGCAAACTCTGCTAATGCATTAAACCCTGAGAAGATCGTTCTAGGCGGCGGTGTGTCTCGTGCAGGCGAGGTATTACTTGCACCGGTAAGAG
ATTATTTCAAACGTTTTGCATTTCCTCGCGTAGCGCAAGGTGCTGAACTAGCAATCGCTACTTTAGGAAACGATGCGGGAATTATTGGAGGAGC
TTGGTTAGTTAAATCTTATTTTGAATAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Part is not RFC10 conform.
- As indicated on the vector map: GCGGCCGCTTCTAG of prefix and TACTAGTAGCGGCCG of suffix are deleted
- Use only after amplification with appropriate primers
Number: [http://parts.igem.org/Part:BBa_K1469004 BBa_K1469004]
Description:
Beta-phosphoglucomutase of Bacillus megaterium. The enzyme catalyses the conversion of glucose 6-phospahate to glucose 1-phosphate.
Sequence:
ATGAAAAAAGAAGAGCTACAAGCGATTATTTTTGATTTAGACGGTGTTATTGCAGATACGGTACATTTATATTATAAAGCAAATAAAAAAGT
AGCAGATCAACTTAACGTTTCATTTTCTGAAGAACTCAATCAGCAGCTGCAGGGGATCAGCCGAGTGAAAACTGTAGAGCTCATTGCTGCCC
AAGGCAATGTACATCTGACAATGGAAAAAAAAGAACAGCTTGCTGATGAAAAGAATAAGCATTATCAAGCATTAATAAAGGAAATGACAAAG
GATCATCTACTTCCAGGAATACAATTGTTGTTAGAGGATTGTAAAAAACAAGGTATTAAAATGGCGATTGCTTCTTCTAGTTCAAATGCAAA
TACTGTAGTGGATGCGTTAGGAATACGTCCTTTTTTTGATTGCATTGTGGATGTGAGAAAAATAAAGAAGGGAAAACCAGACCCAGAAATAT
TTTTGACAGCTGCCCATCAATTAAAAGTAACACCCTATGCATGTGTGGCTATAGAAGATGGAGAGGCGGGTGTAAAAGCTATTAATCAAACA
GATATGGTTTCTATCGGCGTTGGTAAGCATCTGAAGGAAGAGGATATGAATTTTCACGTATATTCAACAGCTGATTTGCATTTAAAGACGAT
CAAAGAGGTTTTCGAAAAAACAAGAAAAGGAACAGATTATAACAGCGTGTATACAAAAAATTAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Part is not RFC10 conform
- As indicated on the vector map: GCGGCCGCTTCTAG of prefix is deleted
- Use only after amplification with appropriate primers
Number: [http://parts.igem.org/Part:BBa_K1469005BBa_K1469005]
Description:
UTP--glucose-1-phosphate uridylyltransferase of Bacillus megaterium. The enzyme catalyses the conversion of glucose 1-phospahate to UDP-glucose 1-phosphate.
Sequence:
ATGACGATAAAAAAGGCAGTTATTCCAGTAGGCGGTTTAGGAACTCGATTTTTGCCAGCGACAAAAGCGCAGCCCAAAGAAATGCTGCCAAT
TGTCGATAAGCCTGCTGTACAATATATTGTAGAAGAAGCTGTGAAATCAGGTATTGAAAGTATTATTTTTATTACAGGACGAAACAAAAAAT
CAATTGAAGACCATTTTGATAAATCAACTGAGTTAGAGCAAATGCTTGAAGAAAAACACAAGTTCCAAATGCTAAAAGAAGTACAAACGATC
TCATCAATGGCCAGCATTCACTATATTCGTCAAAAAGAGCCGTTAGGACTTGGTCATGCTATTCTTTGTGCCGAACAGTTTATTGGAGATGA
GCCGTTTGCCGTTCTTCTTGGAGACGATATTATGACTTCTGAAGAACCTGCGCTCAAACAGATAATCCAAGCGTATGAAACGACTAATCAAT
CAGTTATCGGAGTTCAAAAAGTGGATGTTGAAGAGGTATCAAAATACGGAATCATTCAGCCGAAAAATAAATCTGGCAGCCTTCATGAAGTG
AATGACTTGATTGAAAAACCATCGATAGAACAAGCTCCGTCAACTATTGCCGTTATGGGCCGCTATATTTTAAATCCTTCTATTTTTTCTTA
TCTAAAAACGATTGAAAGAGGGGTAGGAAATGAATTACAATTAACAGATGCACTGGGTGTGGTATGTGAAAAAGAGCGATTGTTTGCTCTTG
AACTGGAAGGACAGCGCTTTGATATTGGAGATAAGGTAGGCTATATGAAAGCGATGGTAGAAGTTGCGTTAAAGCGCAGAGATTTACGTCAA
ACGTTTTTATCGTATTTGGAGGGAATTGTAGAAAAAGAACGAGCAAACAACTTCAGCTAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Part is not RFC10 conform
- As indicated on the vector map: GCGGCCGCTTCTAG of prefix and TACTAGTAGCGGCCG of suffix are deleted
- Use only after amplification with appropriate primers
Number: [http://parts.igem.org/Part:BBa_K1469006 BBa_K1469006]
Description:
Glucose-6-phosphate isomerase of Bacillus megaterium. The enzyme catalyses the conversion of glucose 6-phospahate to fructose 6-phosphate.
Sequence:
ATGACACATGTTCGTTTTGATTATTCAAACGCGTTATCATTTTTTGGTGAACATGAACTTACATACTTACGTGATGCCGTGAAAGTAGCTC
ACCATTCAATTCATGAGAAAACAGGTGCTGGAAACGACTTTTTAGGATGGGTCGACCTTCCTACGGAGTATGACAAAGAAGAATTTGCACG
CATTCAAAAAAGTGCAGAAAAAATTAAATCTGATTCAGATGTATTAATTGTTATCGGTATTGGTGGATCTTACTTAGGTGCTCGTGCTGCT
ATTGAAATGCTTCAGCATTCATTCTATAACACATTATCAAAAGAACAGCGTAAAACTCCGCAAGTTATTTTTGCTGGTAACAACATCAGCT
CAACATACATGAAAGACTTAATGGATGTTTTACAAGATAAAGATTTCTCTATTAATGTTATTTCAAAATCTGGTACAACAACAGAACCGGC
GATTGCTTTCCGTATCTTCCGTAAATTATTAGAAGAGAAATACGGCAAAGAAGAAGCACGCAAGCGTATTTATGCAACAACAGATAAAGCG
CGCGGTGCATTAAAAACATTAGCGGACGAAGAAGGCTATGAAACATTTGTTATTCCAGATGATGTAGGCGGCCGTTATTCAGTATTAACAG
CGGTTGGCTTATTACCAATTGCCGCTGCGGGTCTTGATATCGAACAAATGATGAAAGGGGCAGCTGATGCTCAAGAAGCATTCTCATCATC
AGAGCTAGAAGAGAACCCAGCTTATCAATATGCTGTAGCTCGTAATGTGCTTTATAATAAAGGAAAAACAATTGAAATGCTGATTAACTAT
GAGCCAGCTCTTCAATACTTTGCTGAGTGGTGGAAACAGTTATTTGGCGAAAGTGAAGGAAAAGACCAAAAAGGTATTTACCCTTCATCTG
CTAACTTCTCAACGGATCTTCATTCTTTAGGTCAATATGTACAAGAAGGTCGCCGTGATATTTTTGAAACAATTGTACAAGTTCAAGATTC
TCGTCATGAGCTAACAATTGAAGAAGATGCAGCGGACTTAGATGGATTAAATTACTTAGCTGGTGAAACAGTTGACTTTGTAAATAAAAAA
GCGTTCCAAGGTACAATGCTTGCTCACACAGATGGAGGCGTTCCAAACTTAGTTGTGAACATTCCTGCGCTAGATGAGTATACGTTCGGAT
ATGTTGTATATTTCTTCGAAAAAGCATGTGCAATCAGCGGCTATTTATTAGGCGTAAACCCATTCGATCAGCCGGGAGTAGAAGCATACAA
AGTAAATATGTTTGCTCTTCTTGGAAAACCTGGTTTTGAAGAGAAAAAAGCAGAATTAGAAAAGCGTTTGAAATAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Part is not RFC10 conform
- As indicated on the vector map: TACTAGTAGCGGCCG of suffix is deleted
- Use only after amplification with appropriate primers
Number: [http://parts.igem.org/Part:BBa_K1469007 BBa_K1469007]
Description:
Glucosamine--fructose-6-phosphate aminotransferase (isomerising) of Bacillus megaterium. The enzyme catalyses the conversion of fructose 6-phosphate to glucosamine 6-phosphate.
Sequence:
ATGTGCGGAATTGTAGGATATATTGGAACAGAAGATTCGAAAGAAATTTTATTACGTGGTCTTGAAAAGTTAGAATATCGTGGCTATGACT
CAGCGGGTATTGCAGTTGTTAACAATGAAGGCGTACACGTATTCAAAGAAAAAGGCCGTATTGCAGAATTGCGTCAGGCAGTAGATAACAG
TGTTGTAGCACCAGCAGGTATCGGTCATACGCGTTGGGCAACTCATGGTGTACCAAGCCAAGAAAATGCTCACCCGCATCAAAGTACATCT
GGACGCTTCACACTTGTGCATAACGGTGTAATTGAAAACTATGCTATTTTGAAACGTGAATATTTACAAGATGTAACGTTCAAAAGCGAAA
CGGATACAGAAGTAATCGTTCAGTTAATTCAAAAAATCGCAGCAGAAGGCTTAGATGTAGAAGAAGCGTTCCGTAAGACAGTAAGCTTGCT
TCATGGTTCTTATGCACTAGCTCTTGTTGATAACAAAGATAAAAATACAATCTACGTAGCTAAAAACAAAAGCCCGCTTCTTGTAGGTGTT
GGTGAAGGATTTAACGTTGTAGCGAGTGATGCTATGGCAATGCTTCAAGTAACAGATCAGTACGTAGAATTAATGGATAAAGAAACAGTAA
TTGTTACAAAAGAAGGCGTAACAATTAAAACGCTTGATGGTGAAGTGGTAGAAAGAGCACCTTATACAGCAGAGCTTGATGCAAGTGATAT
TGAAAAAGGTACGTACCCTCACTATATGTTAAAAGAAATTGACGAGCAGCCTTTAGTTGTTCGTAAAATCATTCAAGAGTATCAAAATGAG
AACAATGAACTGCACATTGATGCAGACATTTTAGGAGCAATGGATGAAGCAGACCGCATCTATATCGTAGCATGTGGAACAAGCTATCATG
CAGGCCTAGTAGGTAAACAATTCATTGAAACGTGGGCGAAGGTTCCAGTAGAAGTGCACGTAGCAAGTGAATTTTCTTATAACATGCCGCT
TTTATCTGAAAAGCCATTGTTTATCTTTATTTCTCAAAGTGGAGAAACAGCTGACAGCCGTGCAGTATTAGTTCAAATTAAAGAACTAGGT
TACAAAGCGTTAACAATCACAAACGTTCCGGGATCAACATTATCTCGTGAATCTGATTATACATTATTACTTCATGCAGGTCCGGAAATTG
CTGTAGCATCAACAAAAGCTTATACAGCTCAATTAGCAGTGCTATCAATTCTAGCGGCTGTAACAGCAAAAACACGCGGCATTGAATTAGA
ATTTGATTTAGTTCAAGAGTTAGCAATCGTAGCAAACACAATGGAAGTACTTTGCGACGACAAAGAAGAAATGGAAAGCATTGCTCTTCAA
TTCTTAGCAACAACTCGCAACGCATTCTTTATTGGACGTTCTGTCGATTATTATGTAGGTTTAGAAGGTGCGTTAAAACTAAAAGAAATCT
CTTACATCCAAGCAGAAGGATTTGCTGGAGGAGAGCTTAAGCACGGAACAATCGCTTTAATTGAAAACAATACGCCAATTATTGCATTAGC
AACACAAGAGCATGTAAACTTAAGCATCCGTGGTAACGTAAAAGAAGTAGTAGCACGTGGAGCTAATCCTTGTATCATTTCAATGAAAGGA
TTAGAAACAGAAGAAGATCGCTACGTGATTCCGAAAGTTCACGAAACACTTTCACCACTAGCAGCGGTTATTCCTTTACAATTAATCTCTT
ACTACGCTGCATTGCACCGCGATTGCGATGTAGATAAACCACGTAACTTAGCTAAATCAGTTACTGTAGAATAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Part is not RFC10 conform
- As indicated on the vector map: TACTAGTAGCGGCCG of suffix is deleted
- Use only after amplification with appropriate primers
Number: [http://parts.igem.org/Part:BBa_K1469008 BBa_K1469008]
Description:
Phosphoglucosamine mutase of Bacillus megaterium. The enzyme catalyses the conversion of glucosamine 6-phosphate to glucosamine 1-phosphate.
Sequence:
ATGGGTAAGTATTTTGGAACAGACGGAGTACGAGGAGTTGCCAATAGTGAATTAACTCCTGAGCTTGCTTTTAAAATTGGACGCTTAGGTG
GTTACGTTCTAACAAAAGATTCAGAGCGACCAAAAGTATTAATCGGACGCGATACACGCGTTTCTGGTCATATGTTAGAAGGAGCGCTTGT
TGCAGGATTGCTTTCAATTGGAGCAGAAGTTATGCGACTAGGTGTTATTTCAACTCCAGGGGTAGCGTATTTAACAAAAGCACTTGGCGCA
CAAGCAGGAGTCATGATTTCTGCTTCGCATAACCCAGTTCAAGATAACGGAATTAAATTCTTTGGTCCAGATGGATTTAAACTTTCAGATG
CCCAAGAGAATGAAATTGAAGCGTTAATGGATAGTGACACAGATCAATTACCAAGACCTGTTGGGGGAGACTTAGGTCAGGTAAATGACTA
TTTCGAAGGTGGACAAAAATATCTTCAATACTTAAAACAAACGGTTGATGAAGACTTCTCAGGTATTCATGTGGCATTAGACTGTGCGCAC
GGGGCAACGTCATCGTTAGCAGCACATTTATTTGCAGATCTCGATGCAGATATCTCTACAATGGGGGCATCTCCAAACGGATTGAACATCA
ATGACGGTGTAGGATCTACACATCCGGAAGCATTAAGTGCCTTCTTAAAAGAAAAAGGAGCGGATGTTGGATTAGCATTTGATGGAGACGG
AGATCGTTTAATCGCAATCGACGAAAAAGGCGAGATTGTAGACGGTGATCAAATTATGTTCATTTGTGCAAAATACTTATATGCACAAGGT
CGTTTAAAGAAAAACACGCTTGTTTCTACCGTTATGAGTAACTTAGGCTTCTATAAAGCACTAGAAGCTAGCGGGATTGAAAGTGCACAAA
CTGGTGTAGGAGACCGTTATGTAGTAGAAGAAATGAAAGCTAATCAATATAACCTAGGTGGAGAACAGTCAGGTCATATTATTTTCTTAGA
TTACAACACAACAGGTGACGGTATGCTTTCAGCTATTCAGCTTGTAAACATTATGCAAGCAACGAAAAAGCCTTTATCAGAATTAGCAGCT
GAAATGAAGAAATATCCTCAGCTTCTTGTAAATGTAAAAGTAACAGATAAGCATCATGTTACAGATAACGAGAAGGTAAAAGTGGTTATTG
AAGAAGTGGAAAAAGAAATGAACGGTAATGGCCGAGTACTTGTACGCCCTTCAGGAACAGAGCCGTTAGTACGTGTGATGGTTGAAGCACA
AACACAAGAAGAATGTGAAACTTATGTAACGCGTATCGTAGAAGTAGTAAAAGAAGAAATGGGCTTAGAGTAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Part is not RFC10 conform
- As indicated on the vector map: GCGGCCGCTTCTAG of prefix and TACTAGTAGCGGCCG of suffix are deleted
- Use only after amplification with appropriate primers
Number: [http://parts.igem.org/Part:BBa_K1469009 BBa_K1469009]
Description:
UDP-N-acetylglucosamine pyrophosphorylase of Bacillus megaterium. The enzyme is bifunctional and catalyses the conversion of glucosamine 1-phosphate to N-acetyl glucosamine 1-phosphate as well as the conversion of N-acetyl glucosamine 1-phosphate to UDP-N-acetyl glucosamine, a key precursor molecule for hyaluronic acid production.
Sequence:
ATGTCAAAAAGATATGCAGTCATATTGGCAGCTGGTCAGGGTACACGTATGAAGTCATCTTTATATAAAGTGTTACACCCTGTGTGTGGCAA
ACCGATGGTGCAACACGTAATTGATCAGTTATCACGTTTGGATTTAACAAGTTTAATTACAGTTGTAGGTCACGGTGCCGAAAAAGTGAAAT
CACATGTTGGAGATAAAAGCTTATTTGCTTTACAAGAAGAGCAGTTAGGAACAGCTCATGCAGTAATGCAAGCAGAATCTTTACTTGCACAT
GAAAAAGGTACGACAATTGTTGTATGTGGAGATACTCCGCTTATTACAGCTGAAACAATTGAATCTTTATTAAAGCATCATGAAGCAACAAA
TGCAATGGCAACTATTTTAACAGCTCATGCAGAAGATCCAACAGGATATGGACGCATCATTCGAAATGAACAAGGGTCTGTTGAAAAGATTG
TGGAGCATAAAGATGCCACAGAACAAGAGCGTCTTGTACAGGAAATTAATACTGGTACGTATTGTTTTGATAATGAGAAATTATTTGAGACA
CTTTCAAAAGTTTCAAATGATAATGTACAGGGAGAGTATTATTTACCTGATGTTATCGAAATTCTAAAAAATGAAGGTCAAATTATTTCTGC
ATATCAAACATCCGATTTTGCTGAAACATTGGGCGTTAATGATCGATTTGCTCTGTCTCAAGCAGAAGAAACAATGAAAAAGCGCATCAACA
AGCAGCATATGTTAAATGGAGTGACGATTGTGGATCCTTCTAACACGTACATTTCAGCAGATGCTGTCATTGGCAGAGATACGGTTATTTAT
CCTGGTACGGTTATTCAAGGAACAGTTGTAATTGGAGAGAATTGTGAAGTAGGACCTAACAGTGAAATTAAAGACTGCAAAATAGGTAATAA
TACTTCTATTCGCCATTCTGTTGCTCACGATAGTGAAATTGGTCACGAAGTCACAATTGGCCCATTTGCACATATTCGTCCTCAATCATTAA
TTGGAGATGAAGTGCGCGTAGGAAACTTTGTAGAAATTAAAAAAGCATCTTTTGGCAAAGGTAGCAAAGCTTCTCATTTAAGTTATATTGGA
GACGCAGAGGTCGGAAAAGGTGTAAACTTAGGGTGCGGATCTATTACAGTTAACTACGATGGGAAAAATAAATTTTTAACAAAGATTGAAGA
TGGGGCATTCGTAGGATGTAACTCAAACTTAATTGCACCTGTAACAATTGGTGAAGGCGCCTATGTTGCAGCTGGTTCAACCGTAACGGATG
ATGTGCCAGGAAAAGCATTATCAATTGCTCGTGCGAAACAAGTAAATAAAGAAAACTATGCTGAAAAGCTTGATATTAATAAAAAATCCTAA
Characterisation status:
- Part is user sequenced: no illegal restriction sites, no mutations
- Part is not RFC10 conform
- As indicated on the vector map: GCGGCCGCTTCTAG of prefix and TACTAGTAGCGGCCG of suffix are deleted
- Use only after amplification with appropriate primers
Impressum/Copyright