Synthetic Gene DataBase
 

Synthetic Gene 198


 
  Welcome, Guest!

Field NameNatural GeneSynthetic Gene
SGDB Gene ID184198
GenBank AccessionAF217189DQ289493
GenBank GI745355482880268
Gene NameepoCepoC(SYN)
Gene Length (bp)54995619
SpeciesPolyangium cellulosum (Sorangium cellulosum)Escherichia coli
StrainsSMP44K207-3
CDSatggaagaacaagagtcctccgctatcgcagtcatcggcatgtcgggccgttttccgggg
gcgcgggatctggacgaattctggaggaaccttcgagacggcacggaggccgtgcagcgc
ttctccgagcaggagctcgcggcgtccggagtcgacccagcgctggtgctggacccgaac
tacgtccgggcgggcagcgtgctggaagatgtcgaccggttcgacgctgctttcttcggc
atcagcccgcgcgaggcagagctcatggatccgcagcaccgcatcttcatggaatgcgcc
tgggaggcgctggagaacgccggatacgacccgacagcctacgagggctctatcggcgtg
tacgccggcgccaacatgagctcgtacttgacgtcgaacctccacgagcacccagcgatg
atgcggtggcccggctggtttcagacgttgatcggcaacgacaaggattacctcgcgacc
cacgtctcctacaggctgaatctgagagggccgagcatctccgttcaaactgcctgctct
acctcgctcgtggcggttcacttggcgtgcatgagcctcctggaccgcgagtgcgacatg
gcgctggccggcgggattaccgtccggatcccccatcgagccggctatgtatatgctgag
gggggcatcttctctcccgacggccattgccgggccttcgacgccaaggcgaacggcacg
atcatgggcaacggctgcggggttgtcctcctgaagccgctggaccgggcgctctccgat
ggtgatcccgtccgcgcggtcatccttgggtctgccacaaacaacgacggagcgaggaag
atcgggttcactgcgcccagtgaggtgggccaggcgcaagcgatcatggaggcgctggcg
ctggcaggggtcgaggcccggtccatccaatacatcgagacccacgggaccggcacgctg
ctcggagacgccatcgagacggcggcgttgcggcgggtgttcgatcgcgacgcttcgacc
cggaggtcttgcgcgatcggctccgtgaagaccggcatcggacacctcgaatcggcggct
ggcatcgccggtttgatcaagacggtcttggcgctggagcaccggcagctgccgcccagc
ctgaacttcgagtctcctaacccatcgatcgatttcgcgagcagcccgttctacgtcaat
acctctcttaaggattggaataccggctcgactccgcggcgggccggcgtcagctcgttc
gggatcggcggcaccaacgcccatgtcgtgctggaggaagcacccgcggcgaagcttcca
gccgcggcgccggcgcgctctgccgagctcttcgtcgtctcggccaagagcgcagcggcg
ctggatgccgcggcggcacggctacgagatcatctgcaggcgcaccaggggctttcgttg
ggcgacgtcgccttcagcctggcgacgacgcgcagtcccatggagcaccggctcgcgatg
gcggcaccgtcgcgcgaggcgttgcgagaggggctcgacgcagcggcgcgaggccagacc
ccgccgggcgccgtgcgtggccgctgctccccaggcaacgtgccgaaggtggtcttcgtc
tttcccggccagggctctcagtgggtcggtatgggccgtcagctcctggctgaggaaccc
gtcttccacgcggcgctttcggcgtgcgaccgggccatccaggccgaagctggttggtcg
ctgctcgccgagctcgccgccgacgaagggtcgtcccagatcgagcgcatcgacgtggtg
cagccggtgctgttcgcgctcgcggtggcatttgcggcgctgtggcggtcgtggggtgtc
gggcccgacgtcgtgatcggccacagcatgggcgaggtagccgccgcgcatgtggccggg
gcgctgtcgctcgaggatgcggtggcgatcatctgccggcgcagccggctgctccggcgc
atcagcggtcagggcgagatggcggtgaccgagctgtcgctggccgaggccgaggcagcg
ctccgaggctacgaggatcgggtgagcgtggccgtgagcaacagcccgcgctcgacggtg
ctctcgggcgagccggcagcgatcggcgaggtgctgtcgtccctgaacgcgaagggggtg
ttctgccgtcgggtgaaggtggatgtcgccagccacagcccgcaggtcgacccgctgcgc
gaggacctcttggcagcgctgggcgggctccggccgcgtgcggctgcggtgccgatgcgc
tcgacggtgacgggcgccatggtagcgggcccggagctcggagcgaattactggatgaac
aatctcaggcagcctgtgcgcttcgccgaggtagtccaggcgcagctccaaggcggccac
ggtctgttcgtggagatgagcccgcatccgatcctaacgacttcggtcgaggagatgcgg
cgcgcggcccagcgggcgggcgcagcggtgggctcgctgcggcgagggcaggacgagcgc
ccggcgatgctggaggcgctgggcgcgctgtgggcgcagggctaccctgtaccctggggg
cggctgtttcccgcgggggggcggcgggtaccgctgccgacctatccctggcagcgcgag
cggtactggatcgaagcgccggccaagagcgccgcgggcgatcgccgcggcgtgcgtgcg
ggcggtcacccgctcctcggtgaaatgcagaccctatcaacccagacgagcacgcggctg
tgggagacgacgctggatctcaagcggctgccgtggctcggcgaccaccgggtgcaggga
gcggtcgtgtttccgggcgcggcgtacctggagatggcgatttcgtcgggggccgaggct
ttgggcgatggcccattgcagataaccgacgtggtgctcgccgaggcgctggccttcgcg
ggcgacgcggcggtgttggtccaggtggtgacgacggagcagccgtcgggacggctgcag
ttccagatcgcgagccgggcgccgggcgctggccacgcgtccttccgggtccacgctcgc
ggcgcgttgctccgagtggagcgcaccgaggtcccggctgggcttacgctttccgccgtg
cgcgcacggctccaggccagcatgcccgccgcggccacctacgcggagctgaccgagatg
gggctgcagtacggccctgccttccaggggattgctgagctatggcgcggtgagggcgag
gcgctgggacgggtacgcctgcccgacgcggccggctcggcagcggagtatcggttgcat
cctgcgctgctggacgcgtgcttccaggtcgtcggcagcctcttcgccggcggtggcgag
gcgacgccgtgggtgcccgtggaagtgggctcgctgcggctcttgcagcggccttcgggg
gagctgtggtgccatgcgcgcgtcgtgaaccacgggcgccaaacccccgatcggcagggc
gccgacttttgggtggtcgacagctcgggtgcagtggtcgccgaagtcagcgggctcgtg
gcgcagcggcttccgggaggggtgcgccggcgcgaagaagacgattggttcctggagctc
gagtgggaacccgcagcggtcggcacagccaaggtcaacgcgggccggtggctgctcctc
ggcggcggcggtgggctcggcgccgcgttgcgctcgatgctggaggccggcggccatgcc
gtcgtccatgcggcagagagcaacacgagcgctgccggcgtacgcgcgctcctggcaaag
gcctttgacggccaggctccgacggcggtggtgcacctcggcagcctcgatgggggtggc
gagctcgacccagggctcggggcgcaaggcgcattggacgcgccccggagcgccgacgtc
agtcccgatgccctcgatccggcgctggtacgtggctgtgacagcgtgctctggaccgtg
caggccctggccggcatgggctttcgagacgccccgcgattgtggcttctgacccgcggc
gcacaggccgtcggcgccggcgacgtctccgtgacacaggcaccgctgctggggctgggc
cgcgtcatcgccatggagcacgcggatctgcgctgcgctcgggtcgacctcgatccgacc
cggcccgatggggagctcggtgccctgctggccgagctgctggccgacgacgccgaagcg
gaagtcgcgttgcgcggtggcgagcgatgcgtcgctcggatcgtccgccggcagcccgag
acccggccccgggggaggatcgagagctgcgttccgaccgacgtcaccatccgcgcggac
agcacctaccttgtgaccggcggtctgggtgggctcggtctgagcgtggccggatggctg
gccgagcgcggcgctggtcacctggtgctggtgggccgctccggcgcggcgagcgtggag
caacgggcagccgtcgcggcgctcgaggcccgcggcgcgcgcgtcaccgtggcgaaggca
gatgtcgccgatcgggcgcagctcgagcggatcctccgcgaggttaccacgtcggggatg
ccgctgcggggcgtcgtccatgcggccggcatcttggacgacgggctgctgatgcagcag
actcccgcgcggtttcgtaaggtgatggcgcccaaggtccagggggccttgcacctgcac
gcgttgacgcgcgaagcgccgctttccttcttcgtgctgtacgcttcgggagtagggctc
ttgggctcgccgggccagggcaactacgccgcggccaacacgttcctcgacgctctggcg
caccaccggagggcgcaggggctgccagcgttgagcgtcgactggggcctgttcgcggag
gtgggcatggcggccgcgcaggaagatcgcggcgcgcggctggtctcccgcggaatgcgg
agcctcacccccgacgaggggctgtccgctctggcacggctgctcgaaagcggccgcgtg
caggtgggggtgatgccggtgaacccgcggctgtgggtggagctctaccccgcggcggcg
tcttcgcgaatgttgtcgcgcctggtgacggcgcatcgcgcgagcgccggcgggccagcc
ggggacggggacctgctccgccgcctcgctgctgccgagccgagcgcgcggagcgggctc
ctggagccgctcctccgcgcgcagatctcgcaggtgctgcgcctccccgagggcaagatc
gaggtggacgccccgctcacgagcctgggcatgaactcgctgatggggctcgagctgcgc
aaccgcatcgaggccatgctgggcatcaccgtaccggcaacgctgttgtggacctatccc
acggtggcggcgctgagcgggcatctggcgcgggaggcatgcgaagccgctcctgtggag
tcaccgcacaccaccgccgattctgctgtcgagatcgaggagatgtcgcaggacgatctg
acgcagttgatcgcagcaaaattcaaggcgcttacatga
atggaagaacaagaatccagtgcaattgccgtgattggcatgtcaggtcggtttccaggg
gcccgcgatctggatgagttctggcgcaatctgcgcgacggcaccgaggccgtccagcgc
tttagtgagcaggaactggcggcgtccggcgttgatccggctcttgtgttagatccgaac
tatgtgcgggcaggtagcgttctggaagatgtcgatcgttttgatgccgctttctttggt
atctccccgcgtgaagcggaactgatggacccgcagcaccggatctttatggaatgcgcg
tgggaagcactcgaaaacgccggctatgacccgactgcatacgagggtagcatcggcgtg
tatgcgggggccaacatgagcagttatttaacctcaaatttacatgaacatccggcgatg
atgcgttggccgggttggttccagacgctgatcgggaacgataaagattacttggcaacg
cacgtgtcttaccgtctgaacttgcgtggcccgagtatctccgtccaaactgcgtgctca
acctcgcttgtcgctgttcatttagcttgtatgagcctcctggaccgggaatgcgacatg
gcactggcagggggcatcaccgtccgcatcccgcaccgtgctggttatgtgtacgcggaa
ggcggtattttctcaccagatggtcattgtcgcgcattcgatgccaaggctaatggaacc
attatgggcaatggctgcggcgttgtgctgctgaagccgttagatcgtgcgctgtccgac
ggcgaccctgttcgcgccgtaattctgggcagcgcgaccaataatgacggtgcgcgcaag
attgggtttaccgcgccttcagaggtgggtcaggcgcaagcgatcatggaggcgctggcg
ctggcgggtgttgaggcgcgtagtatccagtacattgaaacacatggcaccggcacactg
ctcggggacgcaatcgaaacggcagccttacgccgcgttttcgatcgcgacgcgtcgact
cgccgctcttgcgccatcggctctgtaaaaaccggcatcggtcatctggaatctgccgct
ggcattgctggtttgattaagaccgtactggcgcttgaacatcgtcagctgccgccttcc
ctcaacttcgaaagcccaaatccgtcgatcgattttgcctcatctccattctacgtgaac
acgtcactgaaagactggaacactggtagcacaccacgccgcgccggggtatcaagcttt
ggtattggcggtaccaacgcccatgtggtgctggaagaagctccggcagccaaattgcca
gctgccgctccagcccgtagcgccgaactgttcgttgtgtcagctaaatcagcagcagcg
ttggatgcagcggcggctcgtctgcgcgatcacctgcaagctcaccagggtttgtccctg
ggcgatgtcgcctttagtctggctactacacgctcccctatggaacatcgtttggcaatg
gcggccccgagtcgggaagcactgcgcgagggtttggatgcggcagcccgtggacaaacg
cctcctggcgcggtccgcggtcgttgttcccctggcaacgtcccgaaagtcgtcttcgtc
tttcctggccagggtagccagtgggtgggtatgggtcgtcagttgttggccgaagaacca
gtttttcatgccgcgctttccgcctgcgatcgtgcaatccaagctgaagctggttggagt
ttattggccgaactggctgccgatgaaggttctagccagatcgaacgtattgacgtggtg
caaccagttctgttcgccttagcagtagcattcgctgccctgtggagatcttggggcgtt
ggtcctgacgtcgtaatcggccatagcatgggtgaggttgcagctgctcacgttgcaggc
gctctgtccctcgaagacgcggtggcaatcatttgtcgccgcagccgtctgctgcggcgt
atttcgggtcagggcgagatggctgttactgaactgagcctcgcggaagcagaagccgcg
ctgcgtggctatgaagaccgtgtctcggtcgcggtgagcaatagcccgcgctctaccgtg
ctgtcgggtgaacctgccgcaatcggggaggttttgtccagcttaaacgcgaagggggta
ttttgtcgtcgcgtgaaagtagatgtggctagccactcaccacaggtagatccattacgt
gaagacctgctggcagcgctgggtggcttacgcccgcgtgcggcggccgtgccgatgcgg
tcaactgtcactggtgcgatggtggcaggcccggaactgggcgctaactactggatgaat
aatctgcgccaaccagttcgcttcgcggaagttgttcaagcgcagctccagggcggtcac
ggtctgtttgtcgaaatgtctccgcatccgattctgaccacctcggtcgaggaaatgcgt
cgggcggcgcaacgcgcaggcgcggcagttggtagcttacgtcgcggccaggatgaacgg
cccgccatgctggaggcgttaggggcgctgtgggcccaaggttatccagttccgtggggg
cgcctttttccggcaggcgggcgccgcgttccgttgccgacttacccttggcagcgtgaa
cgctactggctgcaggcgccagccaaaagcgccgcaggcgatcgtcgcggtgttcgtgca
ggcggccatccgctcttgggcgaaatgcaaaccttatcaacgcaaacgtctacccgcctg
tgggaaaccaccttggatttgaagcgcctgccatggctgggtgatcatcgcgtccagggc
gcagtggtgtttccgggtgcggcctatctggagatggctatttcctcgggtgctgaagcc
ctgggcgatggtccgctacagattacggacgttgttctggcggaggcacttgcgttcgcg
ggcgacgctgcggtactggttcaggtggtgacgacagaacagccgagcgggcgtttacag
tttcagattgcaagccgtgcgccgggtgcgggccacgcgagttttcgtgttcacgcacgc
ggcgctttattacgtgtagagcgcactgaggtgcctgcggggcttacgctttctgcggtc
cgggctcgcttacaggcgtctatgccagccgcagcgacgtatgcggaacttacggagatg
gggctccagtacggtccggcatttcagggcattgccgaactgtggcgcggcgagggggag
gcattgggccgcgtacgtttgccggacgcagcggggagcgccgcggaatatcggctccat
ccagcgctgctggatgcttgctttcaagtggtgggttctttatttgctggcggtggggag
gctaccccgtgggtgccggtggaagttggttctctgcgtctgctgcaacgtccttctggg
gaattatggtgtcacgcacgcgtagttaaccatggccgtcagactccggaccgtcagggt
gccgatttctgggtagtcgacagcagtggcgcggtggtagcggaagtgagtggcctggtg
gcacagcgtttgcctggcggtgtccgccgtcgcgaagaagatgactggtttcttgagctt
gagtgggagccagccgccgtcgggacggctaaggttaatgcgggtcggtggttgctcctg
ggtggcggtggcgggctgggtgctgcacttcgttcgatgctggaagctggcggtcacgcg
gttgtgcatgcggccgagagcaatacatctgcggcgggcgtccgggccctgctagcgaag
gcgttcgatgggcaagctcctacagccgtggttcacctgggctcgctggatggcggtggc
gaacttgacccgggcctgggggcacagggggcgctggatgctcctcgtagtgcagatgtg
tcgccagatgcactggatccggccctggtgcgcggctgcgatagtgtactgtggacggtc
caagcgctggcaggtatgggctttcgcgacgccccgcgtctgtggttgctgactcggggt
gcccaggcggtaggcgccggtgacgtgagtgtgacccaggcaccgctgctcggtttgggt
cgtgttattgccatggaacacgctgacctccgttgtgctcgcgtggatctggatcctacc
cgtccggatggtgaactgggtgcgctgcttgcggaactccttgctgatgatgccgaagcc
gaagttgccttacgtggcggcgagcgctgtgtggctcgcattgttcgccgtcagccggaa
acccgccctcgcggtcgcatcgaaagctgcgtcccaactgatgtgacaatccgtgcagat
agcacctatctggtcaccggtggtcttggcggcttaggcttgtcggttgcgggttggctc
gcggagcgcggtgcaggtcatctggtcctggtaggccgtagcggtgccgcctctgtggag
cagagggctgcggtggcagctttggaagcacgcggggcgcgtgtgaccgtggctaaagct
gacgtagctgatcgcgcccagttagaacgcattttacgggaagtgacgacctcgggcatg
ccgttacgcggcgtcgttcatgccgccgggattctggatgacgggttactgatgcagcaa
acgcccgcacgctttcgtaaagtgatggcgccaaaagttcaaggcgcactccatcttcat
gcactcacgcgcgaggcaccgctgagtttttttgtcctctacgcctccggcgtcggcctg
ttgggttctccgggtcaggggaattatgcggcggccaataccttcttggatgcgctggcg
caccaccgtcgtgctcaggggttaccagccttaagtgtggattggggcctgttcgcggag
gttggtatggctgccgcacaagaagaccggggtgcacgtctggtatcgcgcggcatgcgc
tcgctgaccccggacgaaggtctgagcgctctggctcgtcttcttgaatcgggccgtgtt
caagtgggggtcatgccagtgaaccctcgcctgtgggtggagttgtatccggcggctgcg
agttcacgcatgctgtctcgtctcgtaacagcacatcgtgcatccgctggcggccctgcg
ggcgacggcgatcttctgcgtcgtctggctgcggcggagccttccgcacgttcgggttta
ctggaaccgctccttcgcgcccagatttcacaggtgctgcggctcccagagggcaaaatt
gaggtagatgcgccactgacatccctgggcatgaacagtctcatgggtctggagctgcgg
aaccgtattgaagccatgttgggcattacggttccggcgactcttctttggacgtatccg
accgtagcagcactttcggggcacttagcgcgtgaagcatctagtgctgcgccggtggag
agtccgcatacaaccgcagatagcgcagttgaaatcgaagaaatgtcccaggatgacctg
actcaactgattgccgcgaaatttaaagccctgacggggaattcgagctccgtcgacaag
cttgcggccgcactcgaggtgaagcttgggcccgaacaaaaactcatctcagaagaggat
ctgaatagcgccgtcgaccatcatcatcatcatcattga
5' End
3' End
NotesNo accession number for the natural gene was mentioned in the paper, so the record AF217189 was used.
Expression VectorNAP(BAD)/Myc-HisA (Invitorgen)
Assay MethodsNASDS-PAGE (Coomassie blue staining)
ResultsNot determinedDetectable expression
Protein Functiontype I polyketide synthase
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding MethodE. coli preferred codons were used for the whole gene. Designed by ralph Reid with DNAWorks. To
facilitate cloning, mutations were introduced at the following sites: I884L; E885Q; C1794S; E1795S
Publication Author(s)Mutka SC, Carney JR, Liu Y, Kennedy J.
Corresponding AuthorJonathan Kennedy
Corresponding AddressKosan Biosciences, Inc., 3832 Bay Center Place, Hayward, California 94545.
Publication Year2006
Publication TitleHeterologous Production of Epothilone C and D in Escherichia coli.
AbstractThe epothilones are a family of polyketide natural products that show a high potential as anticancer drugs. They are synthesized by the action of a hybrid nonribosomal peptide synthetase/polyketide synthase in the myxobacterium Sorangium cellulosum. In this work, the genes encoding the entire cluster,epoA, epoB, epoC, epoD, epoE, and epoF, were redesigned and synthesized to allow for expression in Escherichia coli. The expression of the largest of the proteins, EpoD, also required the protein be separated into two polypeptides with compatible module linkers. Using a combination of lowered temperature, chaperone coexpression, and alternative promoters, we succeeded in producing a soluble protein from all genes in the epothilone cluster. The entire synthetic epothilone cluster was then expressed in a strain of E. coli modified to enable polyketide biosynthesis, resulting in the production of epothilones C and D. Furthermore, feeding a thioester of the normal substrate for EpoD to cells expressing the epoD, epoE, and epoF genes also led to the production of epothilones C and D. The design of the synthetic epothilone genes together with E. coli expression provides the ideal platform for both the biochemical investigation of the epothilone PKS and the generation of novel biosynthetic epothilone analogues.
JournalBiochemistry. 2006 Jan 31;45(4):1321-30.. 45(4): 1321-30.
SummaryA cluster of epothilone genes were fully codon-optimized towards E. coli codon preference (with DNAWorks). It appeared that the synthetic epothilone genes were successfully expressed to a detectable level.
Comments
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=505
PubMed ID16430229
Submitter NameWu, Gang
Submitter AddressDepartment of Biological Sciences, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250 USA
Entry ConfirmationNo
 
 

Copyright 2004 the Freeland Bioinformatics Lab, All Rights Reserved. | Contact Us | About this site