LOCUS       Exported               14381 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pMpGWB323 DNA, complete sequence.
ACCESSION   LC057539
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14381)
  AUTHORS   Ishizaki K, Nishihama R, Ueda M, Inoue K, Ishida S, Nishimura Y, 
            Shikanai T, Kohchi T.
  TITLE     Development of Gateway Binary Vector Series with Four Different 
            Selection Markers for the Liverwort Marchantia polymorpha
  JOURNAL   PLoS ONE 10 (9), E0138876 (2015)
  PUBMED    26406247
REFERENCE   2  (bases 1 to 14381)
  AUTHORS   Nishihama R, Ishizaki K, Kohchi T.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-JUN-2015) Contact:Ryuichi Nishihama Kyoto University, 
            Graduate School of Biostudies; Kitashirakawa-oiwake-cho, Sakyo-ku, 
            Kyoto, Kyoto 606-8502, Japan URL :http://www.lif.kyoto-u.ac.jp/e/
REFERENCE   3  (bases 1 to 14381)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14381
                     /organism="Binary vector pMpGWB323"
                     /mol_type="other DNA"
                     /db_xref="taxon:1662964"
     misc_feature    complement(54..78)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     primer_bind     281..297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     complement(323..447)
                     /label=attR1
                     /note="attR1"
     protein_bind    323..447
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        484..514
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             568..1227
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="BAS49954.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            568..1227
                     /gene="cat"
                     /label=cat
     CDS             1569..1874
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAS49955.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1569..1874
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     1915..2039
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(1915..2039)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2053..2070
                     /label=GGSGGS linker
                     /note="GGSGGS linker"
     CDS             2071..4266
                     /codon_start=1
                     /product="three repeats of Citrine with GGSGGS linkers"
                     /label=three repeats of Citrine with GGSGGS linkers
                     /note="region_name: 3xCitrine"
                     /protein_id="BAS49956.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKAGGSGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGE
                     GDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYV
                     QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYI
                     MADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDP
                     NEKRDHMVLLEFVTAAGITLGMDELYKAGGSGGSMVSKGEELFTGVVPILVELDGDVNG
                     HKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDF
                     FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE
                     YNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYL
                     SYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     CDS             2071..2787
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2809..3525
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3547..4263
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      4272..4681
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      4288..4540
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4574..4590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4598..4614
                     /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(4622..4652)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4667..4688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      complement(4763..4952)
                     /regulatory_class="terminator"
                     /note="cauliflower mosaic virus 35S terminator polyA 
                     signal"
     polyA_signal    4773..4948
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(4956..6992)
                     /codon_start=1
                     /gene="mALS"
                     /product="acetolactate synthase"
                     /label=mALS
                     /note="from Marchantia polymorpha; contains triple 
                     amino-acid subustitutions (P207S/R208S/W582L)"
                     /protein_id="BAS49957.1"
                     /translation="MASISGCAAVPLSIASLEKVQPSARSCCKGAKELKVRSGFFSGAA
                     LRGHAYNTSRKTERSLEVKAQAVSEKIQSAVDVSGPRLSSGKSLVAPTPSDFQSRYKDD
                     EPRKGADILVEALEREGIRTVFAYPGGASMEIHQALTRSSSIRNILCRHEQGEIFAAEG
                     YAKSSGRVGVCIATSGPGATNLVTGLADALLDSVPLVAITGQVSSRMIGTDAFQETPIV
                     EVTRSITKHNYLVMNIDDIPRVIREAFFLAASGRPGPVLVDIPKDVQQQMAIPDWSQPM
                     QLKGYLQRLPRPPEPTLIEQIIRLISSSKKPVVYSGGGCNHASEELREFVELTGIPITS
                     TLMGLGAYPESDPLSLKMLGMHGTVYANYAVDQADLLLAFGVRFDDRVTGKLETFASRS
                     SIVHIDIDPAEIGKNKQPHVSMCADVKPALSEINALLKSQKRTFDFSNWVAELEEQKQK
                     WPMKFPVFEDAIAPQSAIQTLYELTGGNAIISTGVGQHQMWAAQWFKYNEPRRWLTSGG
                     LGAMGFGLPSALGAAATQPDIPVIDIDGDGSFVMNVQELATIFVEKLPVKIMILNNQHL
                     GMVVQLEDRFYKANRAHTYLGDPDHEEQIYPDFCTVADGFKVPNARVVKKDELRAAIQT
                     MLDSPGPYLLDVIVPHQEHVLPMIPGGASFKEIITEGDGRTIY"
     gene            complement(4956..6992)
                     /gene="mALS"
                     /label=mALS
     regulatory      complement(7000..7779)
                     /regulatory_class="promoter"
                     /note="double enhancer version of cauliflower mosaic virus 
                     35S promoter"
     promoter        complement(7032..7708)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     misc_feature    complement(7903..7927)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     CDS             8448..9239
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      9485..10073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    10259..10399
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      10743..10937
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11003..12076)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(12505..13134)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"