LOCUS       Exported                8614 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  RNAi vector pGFght-HydA1IR DNA, complete sequence.
ACCESSION   AB909459
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8614)
  AUTHORS   Sato M, Kurahashi A, Nishibori K, Fujimori F.
  TITLE     Development of a transformation system for the edible mushroom 
            Grifola frondosa: demonstrating heterologous gene expression and 
            RNAi-mediated gene silencing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8614)
  AUTHORS   Sato M, Kurahashi A, Nishibori K, Fujimori F.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-FEB-2014) Contact:Masayuki Sato Yukiguni Maitake Co. 
            Ltd., Research and Development; 89 Yokawa, Minamiuonuma, Niigata 
            949-6695, Japan URL :http://www.maitake.co.jp
REFERENCE   3  (bases 1 to 8614)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8614
                     /organism="RNAi vector pGFght-HydA1IR"
                     /mol_type="other DNA"
                     /db_xref="taxon:1461713"
     regulatory      1..1500
                     /regulatory_class="promoter"
                     /gene="Gf.GAPDH"
     gene            1..1500
                     /gene="Gf.GAPDH"
                     /label=Gf.GAPDH
     CDS             1501..2526
                     /codon_start=1
                     /gene="hph"
                     /product="hygromycin B phosphotransferase"
                     /function="hygromycin B resistance"
                     /label=hph
                     /note="derived from pLG-hph"
                     /protein_id="BAQ19371.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     gene            1501..2526
                     /gene="hph"
                     /label=hph
     regulatory      2527..3526
                     /regulatory_class="terminator"
                     /gene="Gf.GAPDH"
     gene            2527..3526
                     /gene="Gf.GAPDH"
                     /label=Gf.GAPDH
     misc_feature    3527..3542
                     /label=multiple cloning site (BamHI-XbaI-SalI) from pUC19
                     /note="multiple cloning site (BamHI-XbaI-SalI) from pUC19"
     gene            3543..5944
                     /gene="Gf.TEF3"
                     /label=Gf.TEF3
     regulatory      3543..4269
                     /regulatory_class="promoter"
                     /gene="Gf.TEF3"
     misc_feature    4270..4591
                     /gene="Gf.TEF3"
                     /label=Gf.HydA1 sense strand sequence
                     /note="Gf.HydA1 sense strand sequence"
     misc_feature    4592..4821
                     /gene="Gf.TEF3"
                     /note="intron of Gf.GAPDH to serve as the spacer for 
                     hairpin formation"
     misc_feature    4822..4829
                     /gene="Gf.TEF3"
                     /label=synthetic sequence for NotI site
                     /note="synthetic sequence for NotI site"
     misc_feature    complement(4830..5151)
                     /label=Gf.HydA1 anti-sense strand sequence
                     /note="Gf.HydA1 anti-sense strand sequence"
     misc_feature    5152..5157
                     /gene="Gf.TEF3"
                     /label=synthetic sequence for SpeI site
                     /note="synthetic sequence for SpeI site"
     regulatory      5158..5944
                     /regulatory_class="terminator"
                     /gene="Gf.TEF3"
     misc_feature    5955..8614
                     /product="pUC19"
                     /label=vector sequences derived from pUC19
                     /note="vector sequences derived from pUC19"
     primer_bind     complement(5972..5988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5996..6012
                     /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(6020..6050)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6065..6086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6374..6962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7133..7993)
                     /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(7994..8098)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8572..8588
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"