LOCUS Exported 6999 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pDEST413CYC1t7, complete sequence.
ACCESSION JX901383
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6999)
AUTHORS Giuraniuc CV, Macpherson M, Saka Y.
TITLE Gateway Vectors for Efficient Artificial Gene Assembly In Vitro and
Expression in Yeast Saccharomyces cerevisiae
JOURNAL PLoS ONE 8 (5), E64419 (2013)
PUBMED 23675537
REFERENCE 2 (bases 1 to 6999)
AUTHORS Giuraniuc C, Saka Y.
TITLE Direct Submission
JOURNAL Submitted (04-OCT-2012) School of Medical Sciences, Institute of
Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen
AB25 2ZD, UK
REFERENCE 3 (bases 1 to 6999)
AUTHORS Giuraniuc C, Saka Y.
TITLE Direct Submission
JOURNAL Submitted (28-JUL-2015) School of Medical Sciences, Institute of
Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen
AB25 2ZD, UK
REFERENCE 4 (bases 1 to 6999)
AUTHORS .
TITLE Direct Submission
COMMENT On Jul 28, 2015 this sequence version replaced JX901383.1.
FEATURES Location/Qualifiers
source 1..6999
/organism="Cloning vector pDEST413CYC1t7"
/mol_type="other DNA"
/db_xref="taxon:1333769"
promoter 317..503
/gene="S. cerevisiae HIS3"
/label=HIS3 promoter
CDS 504..1163
/codon_start=1
/gene="his3"
/product="HIS3"
/label=his3
/note="derived from S. cerevisiae HIS3"
/protein_id="AGL80171.1"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
gene 504..1163
/gene="his3"
/label=his3
rep_origin complement(1423..1878)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin complement(1426..1881)
/direction=LEFT
/label=f1 ori
/note="f1 ori"
primer_bind 2023..2039
/label=M13 -20 primer
/note="M13 -20 primer"
regulatory 2049..2068
/regulatory_class="promoter"
/note="T7 promoter"
promoter 2049..2067
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory complement(2096..2224)
/regulatory_class="terminator"
/note="T7 terminator"
terminator 2121..2168
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 2166..2185
/label=T7 reverse primer
/note="T7 reverse primer"
misc_recomb 2235..2359
/label=attR2 recombination site
/note="attR2 recombination site"
protein_bind 2235..2359
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(2400..2705)
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/protein_id="AGL80172.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene complement(2400..2705)
/gene="ccdB"
/label=ccdB
CDS complement(3047..3706)
/codon_start=1
/gene="CmR"
/product="chloramphenicol resistance protein"
/label=CmR
/protein_id="AGL80173.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene complement(3047..3706)
/gene="CmR"
/label=CmR
promoter complement(3760..3790)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
misc_recomb complement(3815..3939)
/label=attR1 recombination site
/note="attR1 recombination site"
protein_bind complement(3815..3939)
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3960..4206
/regulatory_class="terminator"
/gene="CYC1"
/note="derived from S. cerevisiae CYC1 terminator"
gene 3960..4206
/gene="CYC1"
/label=CYC1
terminator 3961..4207
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
regulatory complement(4224..4243)
/regulatory_class="promoter"
/note="T3 promoter"
promoter complement(4225..4243)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4260..4278)
/label=M13 reverse primer
/note="M13 reverse primer"
primer_bind complement(4264..4280)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4288..4304
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4312..4342)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin complement(4484..5364)
/direction=LEFT
/label=ColE1 ori
/note="ColE1 ori"
rep_origin complement(4666..5254)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5425..6285)
/codon_start=1
/gene="ampR"
/product="beta-lactamase"
/label=ampR
/protein_id="AGL80174.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene complement(5425..6285)
/gene="ampR"
/label=ampR
/note="bla"
promoter complement(6286..6390)
/gene="bla"
/label=AmpR promoter
misc_feature 6427..6930
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
misc_feature 6427..6543
/label=yeast centromere CEN6
/note="yeast centromere CEN6"
rep_origin 6544..6918
/label=yeast replication origin ARSH4
/note="yeast replication origin ARSH4"
LOCUS Exported 6999 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pDEST413CYC1t7, complete sequence.
ACCESSION JX901383
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6999)
AUTHORS Giuraniuc CV, Macpherson M, Saka Y.
TITLE Gateway Vectors for Efficient Artificial Gene Assembly In Vitro and
Expression in Yeast Saccharomyces cerevisiae
JOURNAL PLoS ONE 8 (5), E64419 (2013)
PUBMED 23675537
REFERENCE 2 (bases 1 to 6999)
AUTHORS Giuraniuc C, Saka Y.
TITLE Direct Submission
JOURNAL Submitted (04-OCT-2012) School of Medical Sciences, Institute of
Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen
AB25 2ZD, UK
REFERENCE 3 (bases 1 to 6999)
AUTHORS Giuraniuc C, Saka Y.
TITLE Direct Submission
JOURNAL Submitted (28-JUL-2015) School of Medical Sciences, Institute of
Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen
AB25 2ZD, UK
REFERENCE 4 (bases 1 to 6999)
AUTHORS .
TITLE Direct Submission
COMMENT On Jul 28, 2015 this sequence version replaced JX901383.1.
FEATURES Location/Qualifiers
source 1..6999
/organism="Cloning vector pDEST413CYC1t7"
/mol_type="other DNA"
/db_xref="taxon:1333769"
promoter 317..503
/gene="S. cerevisiae HIS3"
/label=HIS3 promoter
CDS 504..1163
/codon_start=1
/gene="his3"
/product="HIS3"
/label=his3
/note="derived from S. cerevisiae HIS3"
/protein_id="AGL80171.1"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
gene 504..1163
/gene="his3"
/label=his3
rep_origin complement(1423..1878)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin complement(1426..1881)
/direction=LEFT
/label=f1 ori
/note="f1 ori"
primer_bind 2023..2039
/label=M13 -20 primer
/note="M13 -20 primer"
regulatory 2049..2068
/regulatory_class="promoter"
/note="T7 promoter"
promoter 2049..2067
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory complement(2096..2224)
/regulatory_class="terminator"
/note="T7 terminator"
terminator 2121..2168
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 2166..2185
/label=T7 reverse primer
/note="T7 reverse primer"
misc_recomb 2235..2359
/label=attR2 recombination site
/note="attR2 recombination site"
protein_bind 2235..2359
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(2400..2705)
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/protein_id="AGL80172.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene complement(2400..2705)
/gene="ccdB"
/label=ccdB
CDS complement(3047..3706)
/codon_start=1
/gene="CmR"
/product="chloramphenicol resistance protein"
/label=CmR
/protein_id="AGL80173.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene complement(3047..3706)
/gene="CmR"
/label=CmR
promoter complement(3760..3790)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
misc_recomb complement(3815..3939)
/label=attR1 recombination site
/note="attR1 recombination site"
protein_bind complement(3815..3939)
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3960..4206
/regulatory_class="terminator"
/gene="CYC1"
/note="derived from S. cerevisiae CYC1 terminator"
gene 3960..4206
/gene="CYC1"
/label=CYC1
terminator 3961..4207
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
regulatory complement(4224..4243)
/regulatory_class="promoter"
/note="T3 promoter"
promoter complement(4225..4243)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4260..4278)
/label=M13 reverse primer
/note="M13 reverse primer"
primer_bind complement(4264..4280)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4288..4304
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4312..4342)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin complement(4484..5364)
/direction=LEFT
/label=ColE1 ori
/note="ColE1 ori"
rep_origin complement(4666..5254)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5425..6285)
/codon_start=1
/gene="ampR"
/product="beta-lactamase"
/label=ampR
/protein_id="AGL80174.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene complement(5425..6285)
/gene="ampR"
/label=ampR
/note="bla"
promoter complement(6286..6390)
/gene="bla"
/label=AmpR promoter
misc_feature 6427..6930
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
misc_feature 6427..6543
/label=yeast centromere CEN6
/note="yeast centromere CEN6"
rep_origin 6544..6918
/label=yeast replication origin ARSH4
/note="yeast replication origin ARSH4"