LOCUS       Exported               10733 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway binary vector pGWB625 DNA, complete sequence.
ACCESSION   AB543134
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10733)
  AUTHORS   Nakamura S, Mano S, Tanaka Y, Ohnishi M, Nakamori C, Araki M, Niwa 
            T, Nishimura M, Kaminaka H, Nakagawa T, Sato Y, Ishiguro S.
  TITLE     Gateway binary vectors with the bialaphos resistance gene, bar, as a
            selection marker for plant transformation
  JOURNAL   Biosci. Biotechnol. Biochem. 74 (6), 1315-1319 (2010)
  PUBMED    20530878
REFERENCE   2  (bases 1 to 10733)
  AUTHORS   Tanaka Y, Nakamura S, Ishiguro S, Nakagawa T.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JAN-2010) Contact:Tsuyoshi Nakagawa Shimane 
            University, Center for Integrated Research in Science; 1060 
            Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE   3  (bases 1 to 10733)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     constructed using pPZP221.
FEATURES             Location/Qualifiers
     source          1..10733
                     /organism="Gateway binary vector pGWB625"
                     /mol_type="other DNA"
                     /db_xref="taxon:711418"
     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_feature    complement(338..462)
                     /label=attR1
                     /note="attR1"
     protein_bind    338..462
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        499..529
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             583..1242
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="BAI67438.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            583..1242
                     /gene="cat"
                     /label=cat
     CDS             1584..1889
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAI67439.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1584..1889
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    1930..2054
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(1930..2054)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2065..2097
                     /label=T7-epitope
                     /note="T7-epitope"
     CDS             2065..2097
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     regulatory      2103..2512
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      2119..2371
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2405..2421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2429..2445
                     /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(2453..2483)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2498..2519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      2555..2807
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      complement(2557..2823)
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     CDS             complement(2967..3518)
                     /codon_start=1
                     /gene="bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=bar
                     /protein_id="BAI67440.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            complement(2967..3518)
                     /gene="bar"
                     /label=bar
     regulatory      complement(3546..3832)
                     /regulatory_class="promoter"
                     /note="NOS promoter"
     promoter        complement(3570..3749)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    complement(4255..4279)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     CDS             4800..5591
                     /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      5837..6425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    6611..6751
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      7095..7289
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7355..8428)
                     /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(8857..9486)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"