Parts:

Hyaluronic acid production

Number: [http://parts.igem.org/Part:BBa_K1469001 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: [http://parts.igem.org/Part:BBa_K1469002 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: [http://parts.igem.org/Part:BBa_K1469003 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: [http://parts.igem.org/Part:BBa_K1469004 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: [http://parts.igem.org/Part:BBa_K1469005BBa_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: [http://parts.igem.org/Part:BBa_K1469006 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: [http://parts.igem.org/Part:BBa_K1469007 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: [http://parts.igem.org/Part:BBa_K1469008 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: [http://parts.igem.org/Part:BBa_K1469009 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

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