Team:Saarland/Parts

From 2014.igem.org

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Line 7: Line 7:
<br>
<br>
<h1>Parts:</h1>
<h1>Parts:</h1>
-
<br>
 
<h3>Hyaluronic acid production</h3>
<h3>Hyaluronic acid production</h3>
-
 
<h4>Number: [http://parts.igem.org/Part:BBa_K1469001 ''BBa_K1469001'']</h4>  
<h4>Number: [http://parts.igem.org/Part:BBa_K1469001 ''BBa_K1469001'']</h4>  
-
[[File:PSB1C3 Has2.png|thumb|center|300px| <b>Figure 1:</b> vector map of pSB1C3 Has2]]
+
[[File:PSB1C3 Has2 1.png|thumb|center|450px| <b>Figure 1:</b> vector map of pSB1C3 Has2]]
<br>
<br>
<b>Description:</b><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 catalyzes 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>
+
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>
-
<!-- LINK [MODELLING]-->
+
<html>
-
 
+
<a href="https://2014.igem.org/Team:Saarland/Modeling">
-
<b>Sequence:</b> <br>
+
<img src="https://static.igem.org/mediawiki/2014/d/d2/Links_Pfeil.1.png" width="40px">
 +
<b> Modelling </b>
 +
</a>
 +
</html>
 +
<br>
 +
<br>
 +
<b>Sequence:</b>  
<!-- EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469001] -->
<!-- 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>
-
ATGCACTGTGAACGTTTCTTATGTATTTTACGTATTATTGGTACAACATTATTCGGTGTATCTTTATTATTAGGTATTACAGCTGCTTACATTGTA<br>
 
-
GGTTACCAATTCATTCAAACAGATAACTACTACTTCTCTTTCGGTTTATACGGTGCTTTCTTAGCTTCTCACTTAATTATTCAATCTTTATTCGCT<br>
 
-
TTCTTAGAACACCGTAAAATGAAAAAATCTTTAGAAACACCAATTAAATTAAACAAAACAGTAGCTTTATGTATTGCTGCTTACCAAGAAGATCCA<br>
 
-
GATTACTTACGTAAATGTTTACAATCTGTAAAACGTTTAACATACCCAGGTATTAAAGTAGTAATGGTAATTGATGGTAACTCTGATGATGATTTA<br>
 
-
TACATGATGGATATTTTCTCTGAAGTAATGGGTCGTGATAAATCTGCTACATACATTTGGAAAAACAACTTCCACGAAAAAGGTCCAGGTGAAACA<br>
 
-
GATGAATCTCACAAAGAATCTTCTCAACACGTAACACAATTAGTATTATCTTCTAAATCTGTATGTATTATGCAAAAATGGGGTGGTAAACGTGAA<br>
 
-
GTAATGTACACAGCGTTTCGTGCTCTCGGTCGTTCTGTTGATTACGTACAAGTATGTGATTCTGATACAATGTTAGATCCAGCTTCTTCTGTAGAA<br>
 
-
ATGGTAAAAGTATTAGAAGAAGATCCAATGGTAGGTGGTGTAGGTGGTGATGTACAAATTTTAAACAAATACGATTCTTGGATTTCTTTCTTATCT<br>
 
-
TCTGTACGTTACTGGATGGCTTTCAACATTGAACGTGCTTGTCAATCTTACTTCGGTTGTGTACAATGTATTTCTGGTCCATTAGGTATGTACCGT<br>
 
-
AACTCTTTATTACACGAATTTGTAGAAGATTGGTACTCTCAAGAATTTATGGGTAACCAGTGCAGCTTCGGTGATGATCGTCACTTAACAAACCGT<br>
 
-
GTATTATCTTTAGGTTACGCTACAAAATACACAGCTCGTTCTAAATGTTTAACAGAAACACCAATTGAATACTTACGTTGGTTAAACCAACAAACA<br>
 
-
CGTTGGTCTAAATCTTACTTCCGTGAATGGTTATACAACGCTATGTGGTTCCACAAACACCACTTATGGATGACATACGAAGCGGTAATAACAGGC<br>
 
-
TTCTTCCCATTCTTTTTAATTGCTACAGTAATTCAATTATTCTACCGTGGTAAAATTTGGAACATTTTATTATTCTTATTAACAGTACAATTAGTA<br>
 
-
GGTTTAATAAAATCTTCTTTCGCTTCTTGTTTACGTGGTAACATTGTAATGGTATTCATGTCTTTATACTCTGTATTATACATGTCTTCTTTATTA<br>
 
-
CCAGCTAAAATGTTCGCTATTGCTACAATTAACAAAGCTGGTTGGGGTACATCTGGTCGTAAAACAATTGTAGTAAACTTCATTGGTTTAATTCCA<br>
 
-
GTATCTGTATGGTTCACAATTTTATTAGGTGGTGTAATTTTCACAATTTACAAAGAATCTAAAAAACCATTCTCTGAATCTAAACAAACAGTATTA<br>
 
-
ATTGTAGGTACATTATTATACGCTTGTTACTGGGTAATGTTATTAACATTATACGTAGTATTAATAAACAAATGTGGTCGTCGTAAAAAAGGTCAA<br>
 
-
CAATACGATATGGTATTAGATGTATAA<br>
 
-
<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 <!--LINK TO [ RESULTS (Teil 4)] -->
+
*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 optimsation of Hyaluronic acid production</h3>
+
<h3>Pathway engineering for optimisation of hyaluronic acid production</h3>
-
 
+
<h4>Number: [http://parts.igem.org/Part:BBa_K1469002 ''BBa_K1469002'']</h4>
<h4>Number: [http://parts.igem.org/Part:BBa_K1469002 ''BBa_K1469002'']</h4>
-
[[File:PSB1C3 UDP-GlcDH.png|thumb|center|300px| <b>Figure 2:</b> vector map of pSB1C3 UDP-GlcDH]]
+
[[File:PSB1C3 UDP-GlcDH 2.png|thumb|center|450px| <b>Figure 2:</b> vector map of pSB1C3 UDP-GlcDH]]
<br>
<br>
<b>Description:</b><br>
<b>Description:</b><br>
-
UDP-glucose 6-dehydrogenase of<i> Bacillus megaterium</i>. The enzyme catalyzes the NAD dependent oxidation of UDP-glucose to UDP-glucuronic acid, a key precursor molecule for hyaluronic acid production.
+
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>  
<br>  
-
<b>Sequence:</b> <br>  <!--EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469002]-->
+
<b>Sequence:</b> <br>   
-
 
+
<html>
 +
<span style="font-family:Courier New">
ATGACTAATATCACAGTAGCGGGTACTGGTTACGTAGGCTTAGTTACAGGTGTTTGTTTAGCTGAAATAGGGCATGAGGTAACATGTGTAGATAA
ATGACTAATATCACAGTAGCGGGTACTGGTTACGTAGGCTTAGTTACAGGTGTTTGTTTAGCTGAAATAGGGCATGAGGTAACATGTGTAGATAA
TATTCAGGATAAAATTGATATTTTAAATGAAGGAATATCTCCTATCTTTGAACCTGGGCTTGATGAATTAATTATTAAAAACTCTGATGCTGGTA
TATTCAGGATAAAATTGATATTTTAAATGAAGGAATATCTCCTATCTTTGAACCTGGGCTTGATGAATTAATTATTAAAAACTCTGATGCTGGTA
Line 73: Line 85:
ATGATCCTGTAGGTGTAAACAATTTTAAGAAAATATATCCAACACAAGTGTATTATGAAGAAACAATTGAAAAAGCATTAAAAAATGCTGACATT
ATGATCCTGTAGGTGTAAACAATTTTAAGAAAATATATCCAACACAAGTGTATTATGAAGAAACAATTGAAAAAGCATTAAAAAATGCTGACATT
TGTTTTATATTTACAGAGTGGGAAGAAATAAAAAATATGAATTTAAACTTGTTTAACAAACAAATGAAATCTCCTTTTGTCTATGATGGACGTAA
TGTTTTATATTTACAGAGTGGGAAGAAATAAAAAATATGAATTTAAACTTGTTTAACAAACAAATGAAATCTCCTTTTGTCTATGATGGACGTAA
-
TTGTTACAATCTCGATGAAGTTAAAAGAAGTGGTATCAATTATCAATCTATTGGTCGTCCTAATATAAATGGATACAAAGCTAAAATTAAAGCTA
+
TTGTTACAATCTCGATGAAGTTAAAAGAAGTGGTATCAATTATCAATCTATTGGTCGTCCTAATATAAATGGATACAAAGCTAAAATTAAAGCTA<br>
ATGTCTAA<br>
ATGTCTAA<br>
 +
</span>
 +
</html>
<br>
<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 <!--LINK TO [SDS PAGE Teil 4]-->
+
*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.
Line 84: Line 103:
<h4>Number: [http://parts.igem.org/Part:BBa_K1469003 ''BBa_K1469003'']</h4>
<h4>Number: [http://parts.igem.org/Part:BBa_K1469003 ''BBa_K1469003'']</h4>
-
[[File:PSB1C3 Glk.png|thumb|center|300px| <b>Figure 3:</b> vector map of pSB1C3 Glk]]
+
[[File:PSB1C3 Glk 3.png|thumb|center|450px| <b>Figure 3:</b> vector map of pSB1C3 Glk]]
<br>
<br>
<b>Description:</b><br>
<b>Description:</b><br>
-
Glucokinase of <i>Bacillus megaterium</i>. The enzyme catalyzes the phosphorylation reaction of glucose to glucose 6-phospahate.<br>
+
Glucokinase of <i>Bacillus megaterium</i>. The enzyme catalyses the phosphorylation reaction of glucose to glucose 6-phospahate.<br>
-
 
+
-
<b>Sequence: </b> <br> <!--EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469003]-->
+
 +
<b>Sequence: </b>  <br>
 +
<html>
 +
<span style="font-family:Courier New">
ATGGATGACAAATGGTTAGTTGGAGTTGATTTAGGCGGTACAACAATAAAAATGGCCTTTATTAATCATTATGGAGAAATCATTCACAAGTGGA
ATGGATGACAAATGGTTAGTTGGAGTTGATTTAGGCGGTACAACAATAAAAATGGCCTTTATTAATCATTATGGAGAAATCATTCACAAGTGGA
-
AATTAATACGGATGTGAGCGAGCAAGGCCGTAAAATTCCAACGGATATTGCAAAAGCAATTGATAAAAAGTTAAACGATCTTGGAGAAGTAA
+
AATTAATACGGATGTGAGCGAGCAAGGCCGTAAAATTCCAACGGATATTGCAAAAGCAATTGATAAAAAGTTAAACGATCTTGGAGAAGTAAAA
-
AATCAAGGTTAGTAGGAATTGGCATTGGTGCACCGGGGCCTGTCAACTTTGCAAACGGTTCGATTGAAGTAGCTGTCAATTTAGGTTGGGAA
+
TCAAGGTTAGTAGGAATTGGCATTGGTGCACCGGGGCCTGTCAACTTTGCAAACGGTTCGATTGAAGTAGCTGTCAATTTAGGTTGGGAAAAAT
-
AAATTCCCTATAAAAGATATCTTGGAAGTAGAAACTTCTCTTCCTGTTGTAGTAGACAATGATGCAAACATTGCAGCGATTGGAGAAATGTGGA
+
TCCCTATAAAAGATATCTTGGAAGTAGAAACTTCTCTTCCTGTTGTAGTAGACAATGATGCAAACATTGCAGCGATTGGAGAAATGTGGAAGGG
-
AGGGTGCTGGAGACGGAGCAAAAGATTTACTTTGCGTTACGCTTGGCACAGGCGTTGGCGGTGGCGTCATTGCAAACGGTGAAATTGTACAAGG
+
TGCTGGAGACGGAGCAAAAGATTTACTTTGCGTTACGCTTGGCACAGGCGTTGGCGGTGGCGTCATTGCAAACGGTGAAATTGTACAAGGCGTA
-
CGTAAATGGAGCCGCTGGTGAGATCGGGCACATTACTTCTATTCCTGAAGGCGGGGCACCGTGTAACTGCGGTAAAACCGGCTGTTTAGAAACC
+
AATGGAGCCGCTGGTGAGATCGGGCACATTACTTCTATTCCTGAAGGCGGGGCACCGTGTAACTGCGGTAAAACCGGCTGTTTAGAAACCATTG
-
ATTGCTTCAGCAACTGGAATTGTACGTTTAACAATGGAAGAATTAACGGAAACGGACAAACCAAGTGAGCTTCGCACAGTGTTAGAACAAAATG
+
CTTCAGCAACTGGAATTGTACGTTTAACAATGGAAGAATTAACGGAAACGGACAAACCAAGTGAGCTTCGCACAGTGTTAGAACAAAATGGACA
-
GACAAGTTACATCTAAAGATGTATTTGATGCAGCTCGTTCAAAAGACGGGTTAGCTATGCATGTTGTAGATAAAGTTGCTTTTCATTTAGGTCT
+
AGTTACATCTAAAGATGTATTTGATGCAGCTCGTTCAAAAGACGGGTTAGCTATGCATGTTGTAGATAAAGTTGCTTTTCATTTAGGTCTAGCA
-
AGCACTAGCAAACTCTGCTAATGCATTAAACCCTGAGAAGATCGTTCTAGGCGGCGGTGTGTCTCGTGCAGGCGAGGTATTACTTGCACCGGTA
+
CTAGCAAACTCTGCTAATGCATTAAACCCTGAGAAGATCGTTCTAGGCGGCGGTGTGTCTCGTGCAGGCGAGGTATTACTTGCACCGGTAAGAG
-
AGAGATTATTTCAAACGTTTTGCATTTCCTCGCGTAGCGCAAGGTGCTGAACTAGCAATCGCTACTTTAGGAAACGATGCGGGAATTATTGGAG
+
ATTATTTCAAACGTTTTGCATTTCCTCGCGTAGCGCAAGGTGCTGAACTAGCAATCGCTACTTTAGGAAACGATGCGGGAATTATTGGAGGAGC
-
GAGCTTGGTTAGTTAAATCTTATTTTGAATAA<br>
+
TTGGTTAGTTAAATCTTATTTTGAATAA<br>
 +
</span>
 +
</html>
<br>
<br>
<b>Characterisation status:</b>
<b>Characterisation status:</b>
Line 113: Line 135:
<h4>Number: [http://parts.igem.org/Part:BBa_K1469004 ''BBa_K1469004'']</h4>
<h4>Number: [http://parts.igem.org/Part:BBa_K1469004 ''BBa_K1469004'']</h4>
-
[[File:PSB1C3 PgmB.png|thumb|center|300px| <b>Figure 4:</b> vector map of pSB1C3 pgmB]]
+
[[File:PSB1C3 PgmB 4.png|thumb|center|450px| <b>Figure 4:</b> vector map of pSB1C3 pgmB]]
<br>
<br>
<b>Description:</b> <br>
<b>Description:</b> <br>
-
Beta-phosphoglucomutase of <i>Bacillus megaterium</i>. The enzyme catalyzes the conversion of glucose 6-phospahate to glucose 1-phosphate.<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>      <!--EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469004]-->
+
 +
<b>Sequence:</b><br>         
 +
<html>
 +
<span style="font-family:Courier New">
ATGAAAAAAGAAGAGCTACAAGCGATTATTTTTGATTTAGACGGTGTTATTGCAGATACGGTACATTTATATTATAAAGCAAATAAAAAAGT
ATGAAAAAAGAAGAGCTACAAGCGATTATTTTTGATTTAGACGGTGTTATTGCAGATACGGTACATTTATATTATAAAGCAAATAAAAAAGT
AGCAGATCAACTTAACGTTTCATTTTCTGAAGAACTCAATCAGCAGCTGCAGGGGATCAGCCGAGTGAAAACTGTAGAGCTCATTGCTGCCC
AGCAGATCAACTTAACGTTTCATTTTCTGAAGAACTCAATCAGCAGCTGCAGGGGATCAGCCGAGTGAAAACTGTAGAGCTCATTGCTGCCC
Line 129: Line 152:
GATATGGTTTCTATCGGCGTTGGTAAGCATCTGAAGGAAGAGGATATGAATTTTCACGTATATTCAACAGCTGATTTGCATTTAAAGACGAT
GATATGGTTTCTATCGGCGTTGGTAAGCATCTGAAGGAAGAGGATATGAATTTTCACGTATATTCAACAGCTGATTTGCATTTAAAGACGAT
CAAAGAGGTTTTCGAAAAAACAAGAAAAGGAACAGATTATAACAGCGTGTATACAAAAAATTAA<br>
CAAAGAGGTTTTCGAAAAAACAAGAAAAGGAACAGATTATAACAGCGTGTATACAAAAAATTAA<br>
 +
</span>
 +
</html>
<br>
<br>
<b>Characterisation status:</b>
<b>Characterisation status:</b>
Line 139: Line 164:
<h4>Number: [http://parts.igem.org/Part:BBa_K1469005''BBa_K1469005''] </h4>
<h4>Number: [http://parts.igem.org/Part:BBa_K1469005''BBa_K1469005''] </h4>
-
[[File:PSB1C3 GtaB.png|thumb|center|300px| <b>Figure 5:</b> vector map of pSB1C3 gtaB]]
+
[[File:PSB1C3 GtaB 5.png|thumb|center|450px| <b>Figure 5:</b> vector map of pSB1C3 gtaB]]
<br>
<br>
-
 
<b>Description:</b><br>
<b>Description:</b><br>
-
UTP--glucose-1-phosphate uridylyltransferase of <i>Bacillus megaterium</i>. The enzyme catalyzes the conversion of glucose 1-phospahate to UDP-glucose 1-phosphate.<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> <!--EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469005]-->
+
 +
<b>Sequence:</b> <br> 
 +
<html>
 +
<span style="font-family:Courier New">
ATGACGATAAAAAAGGCAGTTATTCCAGTAGGCGGTTTAGGAACTCGATTTTTGCCAGCGACAAAAGCGCAGCCCAAAGAAATGCTGCCAAT
ATGACGATAAAAAAGGCAGTTATTCCAGTAGGCGGTTTAGGAACTCGATTTTTGCCAGCGACAAAAGCGCAGCCCAAAGAAATGCTGCCAAT
TGTCGATAAGCCTGCTGTACAATATATTGTAGAAGAAGCTGTGAAATCAGGTATTGAAAGTATTATTTTTATTACAGGACGAAACAAAAAAT
TGTCGATAAGCCTGCTGTACAATATATTGTAGAAGAAGCTGTGAAATCAGGTATTGAAAGTATTATTTTTATTACAGGACGAAACAAAAAAT
Line 158: Line 183:
AACTGGAAGGACAGCGCTTTGATATTGGAGATAAGGTAGGCTATATGAAAGCGATGGTAGAAGTTGCGTTAAAGCGCAGAGATTTACGTCAA
AACTGGAAGGACAGCGCTTTGATATTGGAGATAAGGTAGGCTATATGAAAGCGATGGTAGAAGTTGCGTTAAAGCGCAGAGATTTACGTCAA
ACGTTTTTATCGTATTTGGAGGGAATTGTAGAAAAAGAACGAGCAAACAACTTCAGCTAA<br>
ACGTTTTTATCGTATTTGGAGGGAATTGTAGAAAAAGAACGAGCAAACAACTTCAGCTAA<br>
 +
</span>
 +
</html>
<br>
<br>
<b>Characterisation status:</b>
<b>Characterisation status:</b>
Line 168: Line 195:
<h4>Number:  [http://parts.igem.org/Part:BBa_K1469006 ''BBa_K1469006'']</h4>
<h4>Number:  [http://parts.igem.org/Part:BBa_K1469006 ''BBa_K1469006'']</h4>
-
[[File:PSB1C3 Pgi.png|thumb|center|300px| <b>Figure 6:</b> vector map of pSB1C3 pgi]]
+
[[File:PSB1C3 Pgi 6.png|thumb|center|450px| <b>Figure 6:</b> vector map of pSB1C3 pgi]]
<br>
<br>
<b>Description:</b><br>
<b>Description:</b><br>
-
Glucose-6-phosphate isomerase of <i>Bacillus megaterium</i>. The enzyme catalyzes the conversion of glucose 6-phospahate to fructose 6-phosphate.<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> <!-- EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469006]-->
+
 +
<b>Sequence:</b> <br>
 +
<html>
 +
<span style="font-family:Courier New">
ATGACACATGTTCGTTTTGATTATTCAAACGCGTTATCATTTTTTGGTGAACATGAACTTACATACTTACGTGATGCCGTGAAAGTAGCTC
ATGACACATGTTCGTTTTGATTATTCAAACGCGTTATCATTTTTTGGTGAACATGAACTTACATACTTACGTGATGCCGTGAAAGTAGCTC
ACCATTCAATTCATGAGAAAACAGGTGCTGGAAACGACTTTTTAGGATGGGTCGACCTTCCTACGGAGTATGACAAAGAAGAATTTGCACG
ACCATTCAATTCATGAGAAAACAGGTGCTGGAAACGACTTTTTAGGATGGGTCGACCTTCCTACGGAGTATGACAAAGAAGAATTTGCACG
Line 191: Line 219:
ATGTTGTATATTTCTTCGAAAAAGCATGTGCAATCAGCGGCTATTTATTAGGCGTAAACCCATTCGATCAGCCGGGAGTAGAAGCATACAA
ATGTTGTATATTTCTTCGAAAAAGCATGTGCAATCAGCGGCTATTTATTAGGCGTAAACCCATTCGATCAGCCGGGAGTAGAAGCATACAA
AGTAAATATGTTTGCTCTTCTTGGAAAACCTGGTTTTGAAGAGAAAAAAGCAGAATTAGAAAAGCGTTTGAAATAA<br>
AGTAAATATGTTTGCTCTTCTTGGAAAACCTGGTTTTGAAGAGAAAAAAGCAGAATTAGAAAAGCGTTTGAAATAA<br>
 +
</span>
 +
</html>
<br>
<br>
<b>Characterisation status:</b>
<b>Characterisation status:</b>
Line 201: Line 231:
<h4>Number: [http://parts.igem.org/Part:BBa_K1469007 ''BBa_K1469007'']</h4>
<h4>Number: [http://parts.igem.org/Part:BBa_K1469007 ''BBa_K1469007'']</h4>
-
<b>Description:</b><br>
+
[[File:PSB1C3 GlmS 7.png|thumb|center|450px| <b>Figure 7:</b> vector map of pSB1C3 glmS]]
-
Glucosamine--fructose-6-phosphate aminotransferase (isomerising) of <i>Bacillus megaterium</i>. The enzyme catalyzes the conversion of fructose 6-phosphate to glucosamine 6-phosphate.<br>
+
<br>
-
<b>Sequence:</b> <br> <!--EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469007]-->
+
<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
ATGTGCGGAATTGTAGGATATATTGGAACAGAAGATTCGAAAGAAATTTTATTACGTGGTCTTGAAAAGTTAGAATATCGTGGCTATGACT
CAGCGGGTATTGCAGTTGTTAACAATGAAGGCGTACACGTATTCAAAGAAAAAGGCCGTATTGCAGAATTGCGTCAGGCAGTAGATAACAG
CAGCGGGTATTGCAGTTGTTAACAATGAAGGCGTACACGTATTCAAAGAAAAAGGCCGTATTGCAGAATTGCGTCAGGCAGTAGATAACAG
Line 226: Line 260:
TTAGAAACAGAAGAAGATCGCTACGTGATTCCGAAAGTTCACGAAACACTTTCACCACTAGCAGCGGTTATTCCTTTACAATTAATCTCTT
TTAGAAACAGAAGAAGATCGCTACGTGATTCCGAAAGTTCACGAAACACTTTCACCACTAGCAGCGGTTATTCCTTTACAATTAATCTCTT
ACTACGCTGCATTGCACCGCGATTGCGATGTAGATAAACCACGTAACTTAGCTAAATCAGTTACTGTAGAATAA<br>
ACTACGCTGCATTGCACCGCGATTGCGATGTAGATAAACCACGTAACTTAGCTAAATCAGTTACTGTAGAATAA<br>
 +
</span>
 +
</html>
<br>
<br>
<b> Characterisation status:</b>
<b> Characterisation status:</b>
Line 236: Line 272:
<h4>Number: [http://parts.igem.org/Part:BBa_K1469008 ''BBa_K1469008'']</h4>
<h4>Number: [http://parts.igem.org/Part:BBa_K1469008 ''BBa_K1469008'']</h4>
-
<b>Description:</b><br>
+
[[File:PSB1C3 GlmM 8.png|thumb|center|450px| <b>Figure 8:</b> vector map of pSB1C3 glmM]]
-
Phosphoglucosamine mutase of <i>Bacillus megaterium</i>. The enzyme catalyzes the conversion of glucosamine 6-phosphate to glucosamine 1-phosphate.<br>
+
<br>
-
<b>Sequence:</b> <br> <!--EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469008]-->
+
<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
ATGGGTAAGTATTTTGGAACAGACGGAGTACGAGGAGTTGCCAATAGTGAATTAACTCCTGAGCTTGCTTTTAAAATTGGACGCTTAGGTG
GTTACGTTCTAACAAAAGATTCAGAGCGACCAAAAGTATTAATCGGACGCGATACACGCGTTTCTGGTCATATGTTAGAAGGAGCGCTTGT
GTTACGTTCTAACAAAAGATTCAGAGCGACCAAAAGTATTAATCGGACGCGATACACGCGTTTCTGGTCATATGTTAGAAGGAGCGCTTGT
Line 256: Line 296:
AAGAAGTGGAAAAAGAAATGAACGGTAATGGCCGAGTACTTGTACGCCCTTCAGGAACAGAGCCGTTAGTACGTGTGATGGTTGAAGCACA
AAGAAGTGGAAAAAGAAATGAACGGTAATGGCCGAGTACTTGTACGCCCTTCAGGAACAGAGCCGTTAGTACGTGTGATGGTTGAAGCACA
AACACAAGAAGAATGTGAAACTTATGTAACGCGTATCGTAGAAGTAGTAAAAGAAGAAATGGGCTTAGAGTAA<br>
AACACAAGAAGAATGTGAAACTTATGTAACGCGTATCGTAGAAGTAGTAAAAGAAGAAATGGGCTTAGAGTAA<br>
 +
</span>
 +
</html>
<br>
<br>
<b>Characterisation status:</b>
<b>Characterisation status:</b>
Line 266: Line 308:
<h4>Number: [http://parts.igem.org/Part:BBa_K1469009 ''BBa_K1469009'']</h4>
<h4>Number: [http://parts.igem.org/Part:BBa_K1469009 ''BBa_K1469009'']</h4>
-
<b>Description:</b><br>
+
[[File:PSB1C3 GcaD neu.png|thumb|center|450px| <b>Figure 9:</b> vector map of pSB1C3 gcaD]]
-
UDP-N-acetylglucosamine pyrophosphorylase of <i>Bacillus megaterium</i>. The enzyme is bifunctional and catalyzes 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>
+
<br>
-
<b>Sequence:</b> <br> <!--EHER LINK ZU [http://parts.igem.org/Part:BBa_K1469009]-->
+
<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
ATGTCAAAAAGATATGCAGTCATATTGGCAGCTGGTCAGGGTACACGTATGAAGTCATCTTTATATAAAGTGTTACACCCTGTGTGTGGCAA
ACCGATGGTGCAACACGTAATTGATCAGTTATCACGTTTGGATTTAACAAGTTTAATTACAGTTGTAGGTCACGGTGCCGAAAAAGTGAAAT
ACCGATGGTGCAACACGTAATTGATCAGTTATCACGTTTGGATTTAACAAGTTTAATTACAGTTGTAGGTCACGGTGCCGAAAAAGTGAAAT
Line 286: Line 332:
TGGGGCATTCGTAGGATGTAACTCAAACTTAATTGCACCTGTAACAATTGGTGAAGGCGCCTATGTTGCAGCTGGTTCAACCGTAACGGATG
TGGGGCATTCGTAGGATGTAACTCAAACTTAATTGCACCTGTAACAATTGGTGAAGGCGCCTATGTTGCAGCTGGTTCAACCGTAACGGATG
ATGTGCCAGGAAAAGCATTATCAATTGCTCGTGCGAAACAAGTAAATAAAGAAAACTATGCTGAAAAGCTTGATATTAATAAAAAATCCTAA<br>
ATGTGCCAGGAAAAGCATTATCAATTGCTCGTGCGAAACAAGTAAATAAAGAAAACTATGCTGAAAAGCTTGATATTAATAAAAAATCCTAA<br>
 +
</span>
 +
</html>
<br>
<br>
<b>Characterisation status:</b>
<b>Characterisation status:</b>

Latest revision as of 12:14, 17 October 2014


Parts:

Hyaluronic acid production

Number: BBa_K1469001

Figure 1: vector map of pSB1C3 Has2


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: BBa_K1469002

Figure 2: vector map of pSB1C3 UDP-GlcDH


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: BBa_K1469003

Figure 3: vector map of pSB1C3 Glk


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: BBa_K1469004

Figure 4: vector map of pSB1C3 pgmB


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: BBa_K1469005

Figure 5: vector map of pSB1C3 gtaB


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: BBa_K1469006

Figure 6: vector map of pSB1C3 pgi


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: BBa_K1469007

Figure 7: vector map of pSB1C3 glmS


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: BBa_K1469008

Figure 8: vector map of pSB1C3 glmM


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: BBa_K1469009

Figure 9: vector map of pSB1C3 gcaD


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