LOCUS       Exported                9809 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNZ4, complete sequence.
ACCESSION   FJ804172
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9809)
  AUTHORS   Zhang N, Magee BB, Magee PT, Cannon R, Schmid J.
  TITLE     A method for mating clinical Candida albicans isolates
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9809)
  AUTHORS   Zhang N, Magee BB, Magee PT, Cannon R, Schmid J.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAR-2009) IMBS, Massey University, Riddet Road, 
            Palmerston North 4410, New Zealand
REFERENCE   3  (bases 1 to 9809)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9809
                     /organism="Cloning vector pNZ4"
                     /mol_type="other DNA"
                     /db_xref="taxon:691219"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     mobile_element  683..1491
                     /mobile_element_type="insertion sequence:TS1"
                     /note="from Candida albicans chromosome 7"
     regulatory      1511..2082
                     /regulatory_class="promoter"
                     /note="PTR from plasmid p99RLU"
     terminator      1644..1831
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        1852..1907
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1858..1876
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    1888..1906
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2083..4189
                     /label=IMH3r ORF plus terminator from plasmid p3408
                     /note="IMH3r ORF plus terminator from plasmid p3408"
     CDS             join(2083..2533,2781..3895)
                     /codon_start=1
                     /product="IMH3r"
                     /label=IMH3r
                     /protein_id="ACY78684.1"
                     /translation="MVFETSKATSYLKDYPKKDGLSVKELIDSTNFGGLTYNDFLILPG
                     LVNFPSSAVSLETKLTKKITLKSPFVSSPMDTVTEENMAIHMALLGGIGIIHHNCTAEE
                     QAEMVRKVKKYENGFINDPVVISPEVTVGEVKKMGEVLGFTSFPVTENGKVGGKLVGII
                     TSRDIQFHEDNKSPVSEVMTKDLVVGKKGISLTDGNELLRSSKKGKLPIVDAEGNLVSL
                     ISRTDLQKNQDYPNASKSFHSKQLLCGAAIGTIDADRERLDKLVEAGLDVVVLDSSNGS
                     SVFQLNMIKWIKEKYPELQVIAGNVVTREQAALLIEAGADALRIGMGSGSICITQEVMA
                     CGRPQGTAVYGVTEFANKFGVPCIADGGIGNIGHITKALALGASCVMMGGLLAGTAETP
                     DDYFYRDGKRLKTYRGMGSIDAMQQTNTNANASTSRYFSEADKVLVAQGVSGSVVDKGS
                     ITKFVPYLYNGLQHSLQDIGIKSIDELRENVDNGEIRFEFRTASAQFEGGVHGLHSYEK
                     RLHN"
     misc_feature    4211..5860
                     /label=tetR-ScHAP4-WH11 from plasmid pCAITHE5
                     /note="tetR-ScHAP4-WH11 from plasmid pCAITHE5"
     CDS             complement(4462..4542)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             complement(5242..5859)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
     regulatory      5860..6387
                     /regulatory_class="promoter"
                     /note="PENO1 from plasmid pCAITHE5"
     mobile_element  6404..7560
                     /mobile_element_type="insertion sequence:TS2"
                     /note="from Candida albicans chromosome 7"
     misc_feature    7573..668
                     /label=plasmid backbone pBSKS(+)
                     /note="plasmid backbone pBSKS(+)"
     primer_bind     complement(7575..7591)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7621..7639)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7660..7676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7684..7700
                     /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)."
     promoter        complement(7708..7738)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7753..7774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8062..8650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8821..9681)
                     /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"
     promoter        complement(9682..9786)
                     /gene="bla"
                     /label=AmpR promoter