Legrand M, Bachellier-Bassi S, Lee KK, Chaudhari Y, Tournu H, Arbogast L, Boyer H, Chauvel M, Cabral V, Maufrais C, Nesseir A, Pelinska I, Permal E, Rossignol T, Walker LA, Zeidler U, Znaidi S, Schoet
pCA-DEST2302 载体载体图谱
pCA-DEST2302 载体载体序列
LOCUS Exported 9308 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCA-DEST2302, complete sequence.
ACCESSION MG188305
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9308)
AUTHORS Legrand M, Bachellier-Bassi S, Lee KK, Chaudhari Y, Tournu H,
Arbogast L, Boyer H, Chauvel M, Cabral V, Maufrais C, Nesseir A,
Pelinska I, Permal E, Rossignol T, Walker LA, Zeidler U, Znaidi S,
Schoeters F, Majgier C, Julien RA, Ma L, Tichit M, Bouchier C, Van
Dyck P, Munro CA, DEnfert C.
TITLE Generating genomic platforms to study C. albicans pathogenesis
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9308)
AUTHORS Bachellier-Bassi S, Arbogast L, Legrand M, Boyer H, Chauvel M,
DEnfert C.
TITLE Direct Submission
JOURNAL Submitted (05-OCT-2017) Mycology, Institut Pasteur, 25 rue du Dr
ROUX, Paris 75015, France
REFERENCE 3 (bases 1 to 9308)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9308
/organism="Cloning vector pCA-DEST2302"
/mol_type="other DNA"
/note="Candida albicans integrative plasmid; Gateway
destination vector; propagate in ccdB resistant Escherichia
coli"
/db_xref="taxon:2081754"
regulatory 1..92
/regulatory_class="promoter"
/note="ampicillin promoter"
promoter 24..128
/gene="bla"
/label=AmpR promoter
CDS 129..989
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
CDS 327..989
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="AUZ17342.1"
/translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
gene 327..989
/gene="bla"
/label=bla
rep_origin 1138..1766
/label=colE1 ori
/note="colE1 ori"
rep_origin 1160..1748
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/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)."
primer_bind 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2171..2189
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
regulatory 2222..3071
/regulatory_class="promoter"
/note="TDH3 promoter"
regulatory 3078..3202
/regulatory_class="attenuator"
/note="Gateway recombination site attR1"
protein_bind 3078..3202
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3206..3262
/regulatory_class="promoter"
/note="lacUV5 promoter"
promoter 3227..3257
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3311..3970
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="AUZ17343.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 3311..3970
/gene="cat"
/label=cat
CDS 4312..4617
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS 4405..4617
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/note="F plasmid toxin"
/protein_id="AUZ17344.1"
/translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
GEEVADLSHRENDIKNAINLMFWGI"
gene 4405..4617
/gene="ccdB"
/label=ccdB
regulatory 4458..4780
/regulatory_class="attenuator"
/note="Gateway recombination site attR2"
protein_bind complement(4658..4782)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 4790..5533
/codon_start=1
/product="enhanced green fluorescent protein"
/label=enhanced green fluorescent protein
/note="eGFP"
/protein_id="AUZ17345.1"
/translation="RTLQGAGAGASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGD
ATYGKLTLKFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQE
RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA
DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
KRDHMVLLEFVTAAGITHGMDELYK"
CDS 4823..5533
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
CTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
TAAGITHGMDELYK"
CDS 5823..6125
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17346.1"
/translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWFDIKAPTTFENRNVGK
TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFH"
gene 5823..6125
/gene="rps1"
/label=rps1
misc_feature 6127..6132
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
misc_feature 6132..6150
/label=I-SceI site for targeted integration
/note="I-SceI site for targeted integration"
misc_feature 6151..6156
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
CDS 6157..6393
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17347.1"
/translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
gene 6157..6393
/gene="rps1"
/label=rps1
primer_bind complement(6681..6697)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 7164..7543
/regulatory_class="promoter"
/note="TEF1 promoter"
promoter 7199..7542
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 7591..8115
/codon_start=1
/gene="sat1"
/product="nourseothricin resistance protein"
/label=sat1
/protein_id="AUZ17348.1"
/translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
WYWFSGAQDDA"
gene 7591..8115
/gene="sat1"
/label=sat1
primer_bind 8270..8301
/standard_name="SP2"
/label=SP2
/note="Illumina paired end sequencing primer 2"
CDS complement(8303..8599)
/codon_start=1
/product="neomycin/kanamycin resistance protein"
/label=neomycin/kanamycin resistance protein
/protein_id="AUZ17349.1"
/translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
primer_bind complement(8618..8650)
/standard_name="SP1"
/label=SP1
/note="Illumina paired end sequencing primer 1"
promoter complement(8673..8691)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8698..8714)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
Legrand M, Bachellier-Bassi S, Lee KK, Chaudhari Y, Tournu H, Arbogast L, Boyer H, Chauvel M, Cabral V, Maufrais C, Nesseir A, Pelinska I, Permal E, Rossignol T, Walker LA, Zeidler U, Znaidi S, Schoet
pCA-DEST2302 载体载体图谱
pCA-DEST2302 载体载体序列
LOCUS Exported 9308 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCA-DEST2302, complete sequence.
ACCESSION MG188305
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9308)
AUTHORS Legrand M, Bachellier-Bassi S, Lee KK, Chaudhari Y, Tournu H,
Arbogast L, Boyer H, Chauvel M, Cabral V, Maufrais C, Nesseir A,
Pelinska I, Permal E, Rossignol T, Walker LA, Zeidler U, Znaidi S,
Schoeters F, Majgier C, Julien RA, Ma L, Tichit M, Bouchier C, Van
Dyck P, Munro CA, DEnfert C.
TITLE Generating genomic platforms to study C. albicans pathogenesis
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9308)
AUTHORS Bachellier-Bassi S, Arbogast L, Legrand M, Boyer H, Chauvel M,
DEnfert C.
TITLE Direct Submission
JOURNAL Submitted (05-OCT-2017) Mycology, Institut Pasteur, 25 rue du Dr
ROUX, Paris 75015, France
REFERENCE 3 (bases 1 to 9308)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9308
/organism="Cloning vector pCA-DEST2302"
/mol_type="other DNA"
/note="Candida albicans integrative plasmid; Gateway
destination vector; propagate in ccdB resistant Escherichia
coli"
/db_xref="taxon:2081754"
regulatory 1..92
/regulatory_class="promoter"
/note="ampicillin promoter"
promoter 24..128
/gene="bla"
/label=AmpR promoter
CDS 129..989
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
CDS 327..989
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="AUZ17342.1"
/translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
gene 327..989
/gene="bla"
/label=bla
rep_origin 1138..1766
/label=colE1 ori
/note="colE1 ori"
rep_origin 1160..1748
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/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)."
primer_bind 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2171..2189
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
regulatory 2222..3071
/regulatory_class="promoter"
/note="TDH3 promoter"
regulatory 3078..3202
/regulatory_class="attenuator"
/note="Gateway recombination site attR1"
protein_bind 3078..3202
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3206..3262
/regulatory_class="promoter"
/note="lacUV5 promoter"
promoter 3227..3257
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3311..3970
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="AUZ17343.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 3311..3970
/gene="cat"
/label=cat
CDS 4312..4617
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS 4405..4617
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/note="F plasmid toxin"
/protein_id="AUZ17344.1"
/translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
GEEVADLSHRENDIKNAINLMFWGI"
gene 4405..4617
/gene="ccdB"
/label=ccdB
regulatory 4458..4780
/regulatory_class="attenuator"
/note="Gateway recombination site attR2"
protein_bind complement(4658..4782)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 4790..5533
/codon_start=1
/product="enhanced green fluorescent protein"
/label=enhanced green fluorescent protein
/note="eGFP"
/protein_id="AUZ17345.1"
/translation="RTLQGAGAGASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGD
ATYGKLTLKFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQE
RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA
DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
KRDHMVLLEFVTAAGITHGMDELYK"
CDS 4823..5533
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
CTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
TAAGITHGMDELYK"
CDS 5823..6125
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17346.1"
/translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWFDIKAPTTFENRNVGK
TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFH"
gene 5823..6125
/gene="rps1"
/label=rps1
misc_feature 6127..6132
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
misc_feature 6132..6150
/label=I-SceI site for targeted integration
/note="I-SceI site for targeted integration"
misc_feature 6151..6156
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
CDS 6157..6393
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17347.1"
/translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
gene 6157..6393
/gene="rps1"
/label=rps1
primer_bind complement(6681..6697)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 7164..7543
/regulatory_class="promoter"
/note="TEF1 promoter"
promoter 7199..7542
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 7591..8115
/codon_start=1
/gene="sat1"
/product="nourseothricin resistance protein"
/label=sat1
/protein_id="AUZ17348.1"
/translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
WYWFSGAQDDA"
gene 7591..8115
/gene="sat1"
/label=sat1
primer_bind 8270..8301
/standard_name="SP2"
/label=SP2
/note="Illumina paired end sequencing primer 2"
CDS complement(8303..8599)
/codon_start=1
/product="neomycin/kanamycin resistance protein"
/label=neomycin/kanamycin resistance protein
/protein_id="AUZ17349.1"
/translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
primer_bind complement(8618..8650)
/standard_name="SP1"
/label=SP1
/note="Illumina paired end sequencing primer 1"
promoter complement(8673..8691)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8698..8714)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"