LOCUS       Exported                5127 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Giardia integration vector pc-CycB-3HA-PAC, complete sequence.
ACCESSION   HQ589229
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5127)
  AUTHORS   Gourguechon S, Cande WZ.
  TITLE     Rapid tagging and integration of genes in Giardia intestinalis
  JOURNAL   Eukaryotic Cell (2010) In press
  PUBMED    21115739
REFERENCE   2  (bases 1 to 5127)
  AUTHORS   Gourguechon S, Cande WZ.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-NOV-2010) Mol and Cell Biology, University of 
            California, Berkeley, 345 LSA, Berkeley, CA 94720-3200, USA
REFERENCE   3  (bases 1 to 5127)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5127
                     /organism="Giardia integration vector pc-CycB-3HA-PAC"
                     /mol_type="other DNA"
                     /db_xref="taxon:931881"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /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"
     CDS             1030..1782
                     /codon_start=1
                     /product="tagged cyclin B"
                     /label=tagged cyclin B
                     /protein_id="ADR65086.1"
                     /translation="KASNRLVGICCLFIAAKYEQSYIPSLRHFVEALTQVDPEFLVNCK
                     CEGNRSGPCYRCSREAIIDTEESILHTLDFQLAMPTILTFLRRYARISEMKNRDRYIAF
                     YISELSCLTTSIFKFKPSEIAAGCIAMSRRLTRKPTWDETLEKYSGYTESDTKKVMAEI
                     STIMKKYACNPKARYIRRKYSQEDRFYGVSLMIETVVTQALTLNTKEAKPGLELKEYPY
                     DVPDYAEYPYDVPDYAEYPYDVPDYAGS"
     misc_feature    1684..1782
                     /label=3HA epitope tag
                     /note="3HA epitope tag"
     CDS             1687..1713
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1717..1743
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1747..1773
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2189..2797
                     /codon_start=1
                     /product="puromycin acetyl transferase"
                     /label=puromycin acetyl transferase
                     /note="puromycin resistance protein"
                     /protein_id="ADR65087.1"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGALE"
     CDS             2189..2788
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     promoter        complement(2939..2957)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2978..2994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3002..3018
                     /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(3026..3056)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3071..3092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3380..3968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4139..4999)
                     /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(5000..5104)
                     /gene="bla"
                     /label=AmpR promoter