LOCUS       Exported                8379 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCK906, complete sequence.
ACCESSION   KM407508
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8379)
  AUTHORS   Kraft C.
  TITLE     A toolbox for the analysis of weak and low abundant molecule 
            interactions in vivo by M-Track in budding yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8379)
  AUTHORS   Brezovich A, Schuschnig M, Ammerer G, Kraft C.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2014) Department of Biochemistry and Cell Biology,
            MFPL, University of Vienna, Dr. Bohr Gasse 9, Vienna 1030, Austria
REFERENCE   3  (bases 1 to 8379)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8379
                     /organism="Cloning vector pCK906"
                     /mol_type="other DNA"
                     /db_xref="taxon:1595838"
     protein_bind    155..176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        191..221
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    229..245
                     /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     253..269
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        290..308
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            345..3570
                     /gene="ATG18"
                     /label=ATG18
     regulatory      345..993
                     /regulatory_class="promoter"
                     /gene="ATG18"
     CDS             994..3570
                     /codon_start=1
                     /gene="ATG18"
                     /product="Atg18"
                     /label=ATG18
                     /protein_id="AJD87343.1"
                     /translation="MSDSSPTINFINFNQTGTCISLGTSKGFKIFNCEPFGKFYSEDSG
                     GYAIVEMLFSTSLLALVGIGDQPALSPRRLRIINTKKHSIICEVTFPTSILSVKMNKSR
                     LVVLLQEQIYIYDINTMRLLHTIETNPNPRGLMAMSPSVANSYLVYPSPPKVINSEIKA
                     HATTNNITLSVGGNTETSFKRDQQDAGHSDISDLDQYSSFTKRDDADPTSSNGGNSSII
                     KNGDVIVFNLETLQPTMVIEAHKGEIAAMAISFDGTLMATASDKGTIIRVFDIETGDKI
                     YQFRRGTYATRIYSISFSEDSQYLAVTGSSKTVHIFKLGHSMSNNKLDSDDSNMEEAAA
                     DDSSLDTTSIDALSDEENPTRLAREPYVDASRKTMGRMIRYSSQKLSRRAARTLGQIFP
                     IKVTSLLESSRHFASLKLPVETNSHVMTISSIGSPIDIDTSEYPELFETGNSASTESYH
                     EPVMKMVPIRVVSSDGYLYNFVMDPERGGDCLILSQYSILMDLQEKISSSGALDYDIPT
                     TASENLYFQGEFGLAQHDEAVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPS
                     QSANLLAEAKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNALIQSLKDDPSQ
                     SANLLAEAKKLNDAQAPKVDANHQLQDDLESRGSPGIPNSARTKQTARKSTGGKAPRKQ
                     LAQIPNSARTKQTARKSTGGKAPRKQLAQIPNSARTKQTARKSTGGKAPRKQLAQIPNS
                     ARTKQTARKSTGGKAPRKQLAQICGRIFYPYDVPDYAGYPYDVPDYAGSYPYDVPDYAA
                     QCVESHHHHHHGDYKDDDDKDYKDDDDKDYKDDDDKDYKDDDDKLQ"
     misc_feature    2554..2574
                     /gene="ATG18"
                     /label=TEV site
                     /note="TEV site"
     CDS             2554..2574
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    2605..2952
                     /gene="ATG18"
                     /label=2x HA
                     /note="2x HA"
     CDS             2605..2778
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2782..2952
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNALIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     misc_feature    3019..3312
                     /gene="ATG18"
                     /label=4x Histone3
                     /note="4x Histone3"
     misc_feature    3337..3426
                     /gene="ATG18"
                     /label=3x HA
                     /note="3x HA"
     CDS             3337..3426
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    3445..3462
                     /gene="ATG18"
                     /label=6x His
                     /note="6x His"
     CDS             3445..3462
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3466..3561
                     /gene="ATG18"
                     /label=4x Flag
                     /note="4x Flag"
     CDS             3466..3537
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             3490..3561
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      3584..3831
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(3850..3868)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3878..3894)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4035..4490
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     gene            complement(4549..5720)
                     /gene="HIS3"
                     /label=HIS3
     CDS             complement(4750..5409)
                     /codon_start=1
                     /gene="HIS3"
                     /product="His3"
                     /label=HIS3
                     /protein_id="AJD87345.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(5410..5596)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    5982..6485
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     rep_origin      5985..6495
                     /label=CEN-ARS
                     /note="CEN-ARS"
     promoter        6522..6626
                     /gene="bla"
                     /label=AmpR promoter
     gene            6557..7487
                     /gene="bla"
                     /label=bla
     regulatory      6557..6585
                     /regulatory_class="promoter"
                     /gene="bla"
     CDS             6627..7487
                     /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             6825..7487
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance protein"
                     /protein_id="AJD87344.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      7658..8246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"