LOCUS       Exported                9113 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCA-DEST2303, complete sequence.
ACCESSION   MG188306
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9113)
  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  JOURNAL   Unpublished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 9113)
  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 9113)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9113
                     /organism="Cloning vector pCA-DEST2303"
                     /mol_type="other DNA"
                     /note="Candida albicans integrative plasmid; Gateway 
                     destination vector; propagate in ccdB resistant Escherichia
                     coli"
                     /db_xref="taxon:2081755"
     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="AUZ17350.1"
                     /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="AUZ17351.1"
                     /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="AUZ17352.1"
                     /translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
                     GEEVADLSHRENDIKNAINLMFWGI"
     gene            4405..4617
                     /gene="ccdB"
                     /label=ccdB
     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"
     regulatory      4658..4780
                     /regulatory_class="attenuator"
                     /note="Gateway recombination site attR2"
     CDS             4793..5338
                     /codon_start=1
                     /product="tandem affinity purification tag protein"
                     /label=tandem affinity purification tag protein
                     /note="TAP tag"
                     /protein_id="AUZ17353.1"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGALDYDIPTTASENLYFQGELK
                     TAALAQHDEAVDNKFNKEQQNAFYEILHLPNLSEEQRNAFIQSLKDDPSQSANLLAEAK
                     KLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEAKK
                     LNGAQAPKVDANSAGKST"
     CDS             4793..4870
                     /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             4898..4918
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4958..5131
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLSEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             5135..5305
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     CDS             5628..5930
                     /codon_start=1
                     /gene="rps1"
                     /product="ribosomal protein 1"
                     /label=rps1
                     /note="RPS1"
                     /protein_id="AUZ17354.1"
                     /translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWFDIKAPTTFENRNVGK
                     TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFH"
     gene            5628..5930
                     /gene="rps1"
                     /label=rps1
     misc_feature    5932..5937
                     /label=StuI site for targeted integration
                     /note="StuI site for targeted integration"
     misc_feature    5937..5955
                     /label=I-SceI site for targeted integration
                     /note="I-SceI site for targeted integration"
     misc_feature    5956..5961
                     /label=StuI site for targeted integration
                     /note="StuI site for targeted integration"
     CDS             5962..6198
                     /codon_start=1
                     /gene="rps1"
                     /product="ribosomal protein 1"
                     /label=rps1
                     /note="RPS1"
                     /protein_id="AUZ17355.1"
                     /translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
                     QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
     gene            5962..6198
                     /gene="rps1"
                     /label=rps1
     primer_bind     complement(6486..6502)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      6969..7348
                     /regulatory_class="promoter"
                     /note="TEF1 promoter"
     promoter        7004..7347
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             7396..7920
                     /codon_start=1
                     /gene="sat1"
                     /product="nourseothricin resistance protein"
                     /label=sat1
                     /protein_id="AUZ17356.1"
                     /translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
                     RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
                     IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
                     WYWFSGAQDDA"
     gene            7396..7920
                     /gene="sat1"
                     /label=sat1
     primer_bind     8074..8105
                     /standard_name="SP2"
                     /label=SP2
                     /note="Illumina paired end sequencing primer 2"
     CDS             complement(8107..8403)
                     /codon_start=1
                     /product="neomycin/kanamycin resistance protein"
                     /label=neomycin/kanamycin resistance protein
                     /protein_id="AUZ17357.1"
                     /translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
                     AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
     primer_bind     complement(8422..8454)
                     /standard_name="SP1"
                     /label=SP1
                     /note="Illumina paired end sequencing primer 1"
     promoter        complement(8478..8496)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8503..8519)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"