LOCUS       Exported                6561 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCN-HHT2-GFP, complete sequence.
ACCESSION   KF040377
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6561)
  AUTHORS   Zordan RE, Ren Y, Pan SJ, Rotondo G, Penas Ade L, Iluore J, Cormack 
            BP.
  TITLE     Expression Plasmids for Use in Candida glabrata
  JOURNAL   G3 (Bethesda) 3 (10), 1675-1686 (2013)
  PUBMED    23934995
REFERENCE   2  (bases 1 to 6561)
  AUTHORS   Zordan RE, Cormack BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-MAY-2013) Molecular Biology and Genetics, Johns 
            Hopkins School of Medicine, 725 N Wolfe St., Hunterian 609, 
            Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 6561)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6561
                     /organism="Cloning vector pCN-HHT2-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1386863"
     regulatory      188..589
                     /regulatory_class="promoter"
                     /note="Saccharomyces cerevisiae TEF1 promoter"
     promoter        188..585
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             599..1171
                     /codon_start=1
                     /gene="NAT1"
                     /product="nourseothricin-resistance protein"
                     /label=NAT1
                     /protein_id="AGT40270.1"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARELGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     gene            599..1171
                     /gene="NAT1"
                     /label=NAT1
     regulatory      1178..1425
                     /regulatory_class="terminator"
                     /note="region downstream of CYC1 orf in Saccharomyces 
                     cerevisiae"
     terminator      1178..1425
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     1625..1641
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1654..2515
                     /regulatory_class="promoter"
                     /note="Candida glabrata HHT2 promoter"
     primer_bind     2516..2532
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2552..3268
                     /codon_start=1
                     /gene="GFP"
                     /product="GFP"
                     /label=GFP
                     /protein_id="AGT40271.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            2552..3268
                     /gene="GFP"
                     /label=GFP
     regulatory      3287..3684
                     /regulatory_class="terminator"
                     /note="region downstream of HIS3 in Candida glabrata BG2"
     primer_bind     complement(3742..3758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3766..3782
                     /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(3790..3820)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3835..3856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4126..4808)
                     /direction=LEFT
                     /label=pBM1(ColE1) origin
                     /note="pBM1(ColE1) origin"
     rep_origin      complement(4144..4732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4903..5763)
                     /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             complement(4903..5565)
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin-resistance protein"
                     /label=bla
                     /protein_id="AGT40272.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     gene            complement(4903..5565)
                     /gene="bla"
                     /label=bla
     promoter        complement(5764..5868)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5900..6491
                     /label=Candida glabrata CEN/ARS
                     /note="Candida glabrata CEN/ARS"