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-DEST1403 载体载体图谱
pCA-DEST1403 载体载体序列
LOCUS Exported 9109 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCA-DEST1403, complete sequence.
ACCESSION MG188292
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9109)
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JOURNAL Unpublished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 9109)
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 9109)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9109
/organism="Cloning vector pCA-DEST1403"
/mol_type="other DNA"
/note="Candida albicans integrative plasmid; Gateway
destination vector; propagate in ccdB resistant Escherichia
coli"
/db_xref="taxon:2081741"
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="AUZ17244.1"
/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"
regulatory 3285..3409
/regulatory_class="attenuator"
/note="Gateway recombination site attR1"
protein_bind 3285..3409
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3413..3469
/regulatory_class="promoter"
/note="lacUV5 promoter"
promoter 3434..3464
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3518..4177
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="AUZ17245.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 3518..4177
/gene="cat"
/label=cat
CDS 4519..4824
/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 4612..4824
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/note="F plasmid toxin"
/protein_id="AUZ17246.1"
/translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
GEEVADLSHRENDIKNAINLMFWGI"
gene 4612..4824
/gene="ccdB"
/label=ccdB
protein_bind complement(4865..4989)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 4865..4987
/regulatory_class="attenuator"
/note="Gateway recombination site attR2"
CDS 5000..5545
/codon_start=1
/product="tandem affinity purification tag protein"
/label=tandem affinity purification tag protein
/note="TAP tag"
/protein_id="AUZ17247.1"
/translation="KRRWKKNFIAVSAANRFKKISSSGALDYDIPTTASENLYFQGELK
TAALAQHDEAVDNKFNKEQQNAFYEILHLPNLSEEQRNAFIQSLKDDPSQSANLLAEAK
KLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEAKK
LNGAQAPKVDANSAGKST"
CDS 5000..5077
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 5105..5125
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 5165..5338
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="VDNKFNKEQQNAFYEILHLPNLSEEQRNAFIQSLKDDPSQSANLL
AEAKKLNDAQAPK"
CDS 5342..5512
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNGAQAPK"
CDS 5835..6137
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17248.1"
/translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWFDIKAPTTFENRNVGK
TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFH"
gene 5835..6137
/gene="rps1"
/label=rps1
misc_feature 6139..6144
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
misc_feature 6144..6162
/label=I-SceI site for targeted integration
/note="I-SceI site for targeted integration"
misc_feature 6163..6168
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
CDS 6169..6405
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17249.1"
/translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
gene 6169..6405
/gene="rps1"
/label=rps1
primer_bind complement(6655..6671)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(6795..7607)
/codon_start=1
/gene="ura3"
/product="Ura3"
/label=ura3
/protein_id="AUZ17250.1"
/translation="MTVNTKTYSERAETHASPVAQRLFRLMELKKTNLCASIDVDTTKE
FLELIDKLGPYVCLIKTHIDIINDFSYESTIEPLLELSRKHQFMIFEDRKFADIGNTVK
KQYIGGVYKISSWADITNAHGVTGNGVVEGLKQGAKETTTNQEPRGLLMLAELSSVGSL
AYGEYSQKTVEIAKSDKEFVIGFIAQRDMGGQEEGFDWLIMTPGVGLDDKGDGLGQQYR
TVDEVVSTGTDIIIVGRGLFGKGRDPDIEGKRYRNAGWNAYLKKTGQL"
gene complement(6795..7607)
/gene="ura3"
/label=ura3
regulatory 7608..8027
/regulatory_class="promoter"
/note="URA3 promoter"
primer_bind 8059..8090
/standard_name="SP2"
/label=SP2
/note="Illumina paired end sequencing primer 2"
CDS complement(8092..8388)
/codon_start=1
/product="neomycin/kanamycin resistance protein"
/label=neomycin/kanamycin resistance protein
/protein_id="AUZ17251.1"
/translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
primer_bind complement(8407..8439)
/standard_name="SP1"
/label=SP1
/note="Illumina paired end sequencing primer 1"
promoter complement(8474..8492)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8499..8515)
/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-DEST1403 载体载体图谱
pCA-DEST1403 载体载体序列
LOCUS Exported 9109 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCA-DEST1403, complete sequence.
ACCESSION MG188292
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9109)
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 9109)
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 9109)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9109
/organism="Cloning vector pCA-DEST1403"
/mol_type="other DNA"
/note="Candida albicans integrative plasmid; Gateway
destination vector; propagate in ccdB resistant Escherichia
coli"
/db_xref="taxon:2081741"
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="AUZ17244.1"
/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"
regulatory 3285..3409
/regulatory_class="attenuator"
/note="Gateway recombination site attR1"
protein_bind 3285..3409
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3413..3469
/regulatory_class="promoter"
/note="lacUV5 promoter"
promoter 3434..3464
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3518..4177
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="AUZ17245.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 3518..4177
/gene="cat"
/label=cat
CDS 4519..4824
/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 4612..4824
/codon_start=1
/gene="ccdB"
/product="CcdB"
/label=ccdB
/note="F plasmid toxin"
/protein_id="AUZ17246.1"
/translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
GEEVADLSHRENDIKNAINLMFWGI"
gene 4612..4824
/gene="ccdB"
/label=ccdB
protein_bind complement(4865..4989)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 4865..4987
/regulatory_class="attenuator"
/note="Gateway recombination site attR2"
CDS 5000..5545
/codon_start=1
/product="tandem affinity purification tag protein"
/label=tandem affinity purification tag protein
/note="TAP tag"
/protein_id="AUZ17247.1"
/translation="KRRWKKNFIAVSAANRFKKISSSGALDYDIPTTASENLYFQGELK
TAALAQHDEAVDNKFNKEQQNAFYEILHLPNLSEEQRNAFIQSLKDDPSQSANLLAEAK
KLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEAKK
LNGAQAPKVDANSAGKST"
CDS 5000..5077
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 5105..5125
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 5165..5338
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="VDNKFNKEQQNAFYEILHLPNLSEEQRNAFIQSLKDDPSQSANLL
AEAKKLNDAQAPK"
CDS 5342..5512
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNGAQAPK"
CDS 5835..6137
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17248.1"
/translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWFDIKAPTTFENRNVGK
TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFH"
gene 5835..6137
/gene="rps1"
/label=rps1
misc_feature 6139..6144
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
misc_feature 6144..6162
/label=I-SceI site for targeted integration
/note="I-SceI site for targeted integration"
misc_feature 6163..6168
/label=StuI site for targeted integration
/note="StuI site for targeted integration"
CDS 6169..6405
/codon_start=1
/gene="rps1"
/product="ribosomal protein 1"
/label=rps1
/note="RPS1"
/protein_id="AUZ17249.1"
/translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
gene 6169..6405
/gene="rps1"
/label=rps1
primer_bind complement(6655..6671)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(6795..7607)
/codon_start=1
/gene="ura3"
/product="Ura3"
/label=ura3
/protein_id="AUZ17250.1"
/translation="MTVNTKTYSERAETHASPVAQRLFRLMELKKTNLCASIDVDTTKE
FLELIDKLGPYVCLIKTHIDIINDFSYESTIEPLLELSRKHQFMIFEDRKFADIGNTVK
KQYIGGVYKISSWADITNAHGVTGNGVVEGLKQGAKETTTNQEPRGLLMLAELSSVGSL
AYGEYSQKTVEIAKSDKEFVIGFIAQRDMGGQEEGFDWLIMTPGVGLDDKGDGLGQQYR
TVDEVVSTGTDIIIVGRGLFGKGRDPDIEGKRYRNAGWNAYLKKTGQL"
gene complement(6795..7607)
/gene="ura3"
/label=ura3
regulatory 7608..8027
/regulatory_class="promoter"
/note="URA3 promoter"
primer_bind 8059..8090
/standard_name="SP2"
/label=SP2
/note="Illumina paired end sequencing primer 2"
CDS complement(8092..8388)
/codon_start=1
/product="neomycin/kanamycin resistance protein"
/label=neomycin/kanamycin resistance protein
/protein_id="AUZ17251.1"
/translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
primer_bind complement(8407..8439)
/standard_name="SP1"
/label=SP1
/note="Illumina paired end sequencing primer 1"
promoter complement(8474..8492)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8499..8515)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"