LOCUS       Exported                6655 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pB-TUB_DEST_GFP DNA, complete sequence.
ACCESSION   LC136879
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  AUTHORS   Irie T, Sakamoto T.
  TITLE     Construction of selectable maker for Pleurotus ostreatus 
            transformation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6655)
  AUTHORS   Irie T, Sakamoto T.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2016) Contact:Toshikazu Irie University of Shiga 
            Prefecture, School of Environmental Science; 2500, Hassaka-cho, 
            Hikone, Shiga Prefecture 522-8533, Japan URL :http://www.usp.ac.jp/
REFERENCE   3  (bases 1 to 6655)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6655
                     /organism="Expression vector pB-TUB_DEST_GFP"
                     /mol_type="other DNA"
                     /label=for Pleurotus ostreatus transformation
                     /note="for Pleurotus ostreatus transformation"
                     /db_xref="taxon:1840722"
     regulatory      16..1699
                     /regulatory_class="promoter"
                     /gene="b-tub"
                     /note="beta-tubulin gene in Pleurotus ostreatus"
     gene            16..1699
                     /gene="b-tub"
                     /label=b-tub
     misc_recomb     1706..1730
                     /label=attB1
                     /note="attB1"
     protein_bind    1706..1730
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1751..2770
                     /codon_start=1
                     /gene="hph"
                     /product="hygromycin B phosphotransferase"
                     /label=hph
                     /protein_id="BAU94258.1"
                     /translation="MPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     gene            1751..2770
                     /gene="hph"
                     /label=hph
     misc_recomb     2783..2811
                     /label=attB2
                     /note="attB2"
     protein_bind    complement(2787..2811)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2816..3535
                     /codon_start=1
                     /gene="GFP"
                     /product="green fluorescent protein"
                     /label=GFP
                     /protein_id="BAU94259.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            2816..3535
                     /gene="GFP"
                     /label=GFP
     regulatory      3542..3730
                     /regulatory_class="terminator"
                     /gene="sdi1"
                     /note="Ip subunit gene in Pleurotus ostreatus"
     gene            3542..3730
                     /gene="sdi1"
                     /label=sdi1
     promoter        complement(3804..3822)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3840..3856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3864..3880
                     /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(3888..3918)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3933..3954
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4242..4830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5001..5861)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /protein_id="BAU94260.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5001..5861)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(5862..5966)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6044..6499)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"