LOCUS       Exported                9335 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCA-DEST2430, complete sequence.
ACCESSION   MG188316
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9335)
  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  JOURNAL   Unpublished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 9335)
  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 9335)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9335
                     /organism="Cloning vector pCA-DEST2430"
                     /mol_type="other DNA"
                     /note="Candida albicans integrative plasmid; Gateway 
                     destination vector; propagate in ccdB resistant Escherichia
                     coli"
                     /db_xref="taxon:2081765"
     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="AUZ17426.1"
                     /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             3295..3840
                     /codon_start=1
                     /product="tandem affinity purification tag protein"
                     /label=tandem affinity purification tag protein
                     /note="TAP tag"
                     /protein_id="AUZ17427.1"
                     /translation="MKRRWKKNFIAVSAANRFKKISSSGALDYDIPTTASENLYFQGEL
                     KTAALAQHDEAVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEA
                     KKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEAK
                     KLNGAQAPKVDANSAGKST"
     CDS             3298..3375
                     /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             3403..3423
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3463..3636
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             3640..3810
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     regulatory      3844..3968
                     /regulatory_class="attenuator"
                     /note="Gateway recombination site attR1"
     protein_bind    3844..3968
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3979..4028
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     promoter        3993..4023
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4077..4736
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AUZ17428.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            4077..4736
                     /gene="cat"
                     /label=cat
     CDS             5078..5383
                     /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             5171..5383
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="F plasmid toxin"
                     /protein_id="AUZ17429.1"
                     /translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
                     GEEVADLSHRENDIKNAINLMFWGI"
     gene            5171..5383
                     /gene="ccdB"
                     /label=ccdB
     protein_bind    complement(5424..5548)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      5424..5546
                     /regulatory_class="attenuator"
                     /note="Gateway recombination site attR2"
     CDS             5850..6152
                     /codon_start=1
                     /gene="rps1"
                     /product="ribosomal protein 1"
                     /label=rps1
                     /note="RPS1"
                     /protein_id="AUZ17430.1"
                     /translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWFDIKAPTTFENRNVGK
                     TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFH"
     gene            5850..6152
                     /gene="rps1"
                     /label=rps1
     misc_feature    6154..6159
                     /label=StuI site for targeted integration
                     /note="StuI site for targeted integration"
     misc_feature    6159..6177
                     /label=I-SceI site for targeted integration
                     /note="I-SceI site for targeted integration"
     misc_feature    6178..6183
                     /label=StuI site for targeted integration
                     /note="StuI site for targeted integration"
     CDS             6184..6420
                     /codon_start=1
                     /gene="rps1"
                     /product="ribosomal protein 1"
                     /label=rps1
                     /note="RPS1"
                     /protein_id="AUZ17431.1"
                     /translation="HGLDFTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKR
                     QPNQIKETTYAQSSKLREVRKKMIEIMQREVSN"
     gene            6184..6420
                     /gene="rps1"
                     /label=rps1
     primer_bind     complement(6708..6724)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      7191..7570
                     /regulatory_class="promoter"
                     /note="TEF1 promoter"
     promoter        7226..7569
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             7618..8142
                     /codon_start=1
                     /gene="sat1"
                     /product="nourseothricin resistance protein"
                     /label=sat1
                     /protein_id="AUZ17432.1"
                     /translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
                     RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
                     IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
                     WYWFSGAQDDA"
     gene            7618..8142
                     /gene="sat1"
                     /label=sat1
     primer_bind     8297..8328
                     /standard_name="SP2"
                     /label=SP2
                     /note="Illumina paired end sequencing primer 2"
     CDS             complement(8330..8626)
                     /codon_start=1
                     /product="neomycin/kanamycin resistance protein"
                     /label=neomycin/kanamycin resistance protein
                     /protein_id="AUZ17433.1"
                     /translation="ARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAP
                     AEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ"
     primer_bind     8645..8677
                     /standard_name="SP1"
                     /label=SP1
                     /note="Illumina paired end sequencing primer 1"
     promoter        complement(8700..8718)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8725..8741)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"