LOCUS       Exported                9320 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCA-DEST2403, complete sequence.
ACCESSION   MG188313
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9320)
  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  JOURNAL   Unpublished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 9320)
  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 9320)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9320
                     /organism="Cloning vector pCA-DEST2403"
                     /mol_type="other DNA"
                     /note="Candida albicans integrative plasmid; Gateway 
                     destination vector; propagate in ccdB resistant Escherichia
                     coli"
                     /db_xref="taxon:2081762"
     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="AUZ17403.1"
                     /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="AUZ17404.1"
                     /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="AUZ17405.1"
                     /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="AUZ17406.1"
                     /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="AUZ17407.1"
                     /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="AUZ17408.1"
                     /translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
                     QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
     gene            6169..6405
                     /gene="rps1"
                     /label=rps1
     primer_bind     complement(6693..6709)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      7176..7555
                     /regulatory_class="promoter"
                     /note="TEF1 promoter"
     promoter        7211..7554
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             7603..8127
                     /codon_start=1
                     /gene="sat1"
                     /product="nourseothricin resistance protein"
                     /label=sat1
                     /protein_id="AUZ17409.1"
                     /translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
                     RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
                     IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
                     WYWFSGAQDDA"
     gene            7603..8127
                     /gene="sat1"
                     /label=sat1
     primer_bind     8282..8313
                     /standard_name="SP2"
                     /label=SP2
                     /note="Illumina paired end sequencing primer 2"
     CDS             complement(8315..8611)
                     /codon_start=1
                     /product="neomycin/kanamycin resistance protein"
                     /label=neomycin/kanamycin resistance protein
                     /protein_id="AUZ17410.1"
                     /translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
                     AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
     primer_bind     8630..8662
                     /standard_name="SP1"
                     /label=SP1
                     /note="Illumina paired end sequencing primer 1"
     promoter        complement(8685..8703)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8710..8726)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"