LOCUS       Exported               12709 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway binary vector pGWB729 DNA, complete sequence.
ACCESSION   AB608295
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12709)
  AUTHORS   Tanaka Y, Nakamura S, Kawamukai M, Koizumi N, Nakagawa T.
  TITLE     Development of a series of gateway binary vectors possessing a 
            tunicamycin resistance gene as a marker for the transformation of 
            Arabidopsis thaliana
  JOURNAL   Biosci. Biotechnol. Biochem. 75 (4), 804-807 (2011)
  PUBMED    21512216
REFERENCE   2  (bases 1 to 12709)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-JAN-2011) Contact:Tsuyoshi Nakagawa Shimane 
            University, Center for Integrated Research in Science; 1060 
            Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE   3  (bases 1 to 12709)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     constructed using pPZP221.
FEATURES             Location/Qualifiers
     source          1..12709
                     /organism="Gateway binary vector pGWB729"
                     /mol_type="other DNA"
                     /db_xref="taxon:944731"
     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"
     regulatory      325..1159
                     /regulatory_class="promoter"
                     /note="35S promoter"
     promoter        812..1157
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    complement(1179..1303)
                     /label=attR1
                     /note="attR1"
     protein_bind    1179..1303
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1340..1370
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1424..2083
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="BAJ61242.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            1424..2083
                     /gene="cat"
                     /label=cat
     CDS             2425..2730
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAJ61243.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            2425..2730
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    2771..2895
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(2771..2895)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2906..3451
                     /label=TAP
                     /note="TAP"
     CDS             2906..2983
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             3011..3031
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3071..3244
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             3248..3418
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     regulatory      3457..3866
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      3473..3725
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3759..3775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3783..3799
                     /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(3807..3837)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3852..3873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      complement(3902..4161)
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      3909..4161
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4199..5494)
                     /codon_start=1
                     /gene="gpt"
                     /product="UDP-N-acetylglucosamine: dolichol phosphate 
                     N-acetylglucosamine-1-P transferase"
                     /label=gpt
                     /protein_id="BAJ61244.1"
                     /translation="MAARKRASSISIPNKPDPSEPNSAPSEQKMTRKTVSASGEEFRLA
                     PPKLGVIFVISTLLCSLYLYLLCFHYKVDNELKRSILINAGLSLVGFFVTLKMIPVTAR
                     YVLRRNMFGFDINKRGTPQGDIKVPESLGIVVGIVFLIVAIIFQYFNFTEDSNWLVEYN
                     AALASICFMILLGFVDDVLDVPWRVKLVLPSFATLPLLMAYAGHTTIVIPKPLVAYIGL
                     EVLNLGRIYKLYMGLLAVFCTNSINIHAGLNGLEIGQTVVIAAAILIHNVMQIGASVDP
                     EYHQAHAFSIFLTQPLMATSLAMLAYNWYPSSVFVGDTYTVFAGMTMAVVGILGHFSET
                     LLIFFLPQVLNLLLSLPQLAGIVKCPRHRLPRYDPATGLLTGTKDGTLVNVYLRLFGPK
                     SEKSLCIHLLVFQALACAFCFILRHFLAGWYK"
     gene            complement(4199..5494)
                     /gene="gpt"
                     /label=gpt
     regulatory      complement(5522..5808)
                     /regulatory_class="promoter"
                     /note="NOS promoter"
     promoter        complement(5546..5725)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    complement(6231..6255)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     CDS             6776..7567
                     /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      7813..8401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    8587..8727
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      9071..9265
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9331..10404)
                     /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(10833..11462)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"