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-DEST2420 载体载体图谱
pCA-DEST2420 载体载体序列
LOCUS Exported 9516 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCA-DEST2420, complete sequence.
ACCESSION MG188315
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9516)
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JOURNAL Unpublished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 9516)
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 9516)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9516
/organism="Cloning vector pCA-DEST2420"
/mol_type="other DNA"
/note="Candida albicans integrative plasmid; Gateway
destination vector; propagate in ccdB resistant Escherichia
coli"
/db_xref="taxon:2081764"
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="AUZ17418.1"
/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 2736..3233
/regulatory_class="promoter"
/note="ACT1 promoter"
CDS 3284..3955
/codon_start=1
/product="enhanced green fluorescent protein"
/label=enhanced green fluorescent protein
/note="eGFP"
/protein_id="AUZ17419.1"
/translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EF"
CDS 3293..4000
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
CTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
TAAGITHGMDELYK"
regulatory 4025..4149
/regulatory_class="attenuator"
/note="Gateway recombination site attR1"
protein_bind 4025..4149
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 4160..4209
/regulatory_class="promoter"
/note="lacUV5 promoter"
promoter 4174..4204
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 4258..4917
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="AUZ17420.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 4258..4917
/gene="cat"
/label=cat
CDS 5259..5564
/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 5352..5564
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/note="F plasmid toxin"
/protein_id="AUZ17421.1"
/translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
GEEVADLSHRENDIKNAINLMFWGI"
gene 5352..5564
/gene="ccdB"
/label=ccdB
protein_bind complement(5605..5729)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 5605..5727
/regulatory_class="attenuator"
/note="Gateway recombination site attR2"
CDS 6031..6333
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17422.1"
/translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWFDIKAPTTFENRNVGK
TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFH"
gene 6031..6333
/gene="rps1"
/label=rps1
misc_feature 6335..6340
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
misc_feature 6340..6358
/label=I-SceI site for targeted integration
/note="I-SceI site for targeted integration"
misc_feature 6359..6364
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
CDS 6365..6601
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17423.1"
/translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
gene 6365..6601
/gene="rps1"
/label=rps1
primer_bind complement(6889..6905)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 7372..7751
/regulatory_class="promoter"
/note="TEF1 promoter"
promoter 7407..7750
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 7799..8323
/codon_start=1
/gene="sat1"
/product="nourseothricin resistance protein"
/label=sat1
/protein_id="AUZ17424.1"
/translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
WYWFSGAQDDA"
gene 7799..8323
/gene="sat1"
/label=sat1
primer_bind 8478..8509
/standard_name="SP2"
/label=SP2
/note="Illumina paired end sequencing primer 2"
CDS complement(8511..8807)
/codon_start=1
/product="neomycin/kanamycin resistance protein"
/label=neomycin/kanamycin resistance protein
/protein_id="AUZ17425.1"
/translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
primer_bind 8826..8858
/standard_name="SP1"
/label=SP1
/note="Illumina paired end sequencing primer 1"
promoter complement(8881..8899)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8906..8922)
/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-DEST2420 载体载体图谱
pCA-DEST2420 载体载体序列
LOCUS Exported 9516 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCA-DEST2420, complete sequence.
ACCESSION MG188315
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9516)
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 9516)
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 9516)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9516
/organism="Cloning vector pCA-DEST2420"
/mol_type="other DNA"
/note="Candida albicans integrative plasmid; Gateway
destination vector; propagate in ccdB resistant Escherichia
coli"
/db_xref="taxon:2081764"
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="AUZ17418.1"
/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 2736..3233
/regulatory_class="promoter"
/note="ACT1 promoter"
CDS 3284..3955
/codon_start=1
/product="enhanced green fluorescent protein"
/label=enhanced green fluorescent protein
/note="eGFP"
/protein_id="AUZ17419.1"
/translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EF"
CDS 3293..4000
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
CTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
TAAGITHGMDELYK"
regulatory 4025..4149
/regulatory_class="attenuator"
/note="Gateway recombination site attR1"
protein_bind 4025..4149
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 4160..4209
/regulatory_class="promoter"
/note="lacUV5 promoter"
promoter 4174..4204
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 4258..4917
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="AUZ17420.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 4258..4917
/gene="cat"
/label=cat
CDS 5259..5564
/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 5352..5564
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/note="F plasmid toxin"
/protein_id="AUZ17421.1"
/translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
GEEVADLSHRENDIKNAINLMFWGI"
gene 5352..5564
/gene="ccdB"
/label=ccdB
protein_bind complement(5605..5729)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 5605..5727
/regulatory_class="attenuator"
/note="Gateway recombination site attR2"
CDS 6031..6333
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17422.1"
/translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWFDIKAPTTFENRNVGK
TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFH"
gene 6031..6333
/gene="rps1"
/label=rps1
misc_feature 6335..6340
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
misc_feature 6340..6358
/label=I-SceI site for targeted integration
/note="I-SceI site for targeted integration"
misc_feature 6359..6364
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
CDS 6365..6601
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17423.1"
/translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
gene 6365..6601
/gene="rps1"
/label=rps1
primer_bind complement(6889..6905)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 7372..7751
/regulatory_class="promoter"
/note="TEF1 promoter"
promoter 7407..7750
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 7799..8323
/codon_start=1
/gene="sat1"
/product="nourseothricin resistance protein"
/label=sat1
/protein_id="AUZ17424.1"
/translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
WYWFSGAQDDA"
gene 7799..8323
/gene="sat1"
/label=sat1
primer_bind 8478..8509
/standard_name="SP2"
/label=SP2
/note="Illumina paired end sequencing primer 2"
CDS complement(8511..8807)
/codon_start=1
/product="neomycin/kanamycin resistance protein"
/label=neomycin/kanamycin resistance protein
/protein_id="AUZ17425.1"
/translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
primer_bind 8826..8858
/standard_name="SP1"
/label=SP1
/note="Illumina paired end sequencing primer 1"
promoter complement(8881..8899)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8906..8922)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"