LOCUS       Exported               12254 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway binary vector pGWB661 DNA, complete sequence.
ACCESSION   AB543155
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12254)
  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 12254)
  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 12254)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     constructed using pPZP221.
FEATURES             Location/Qualifiers
     source          1..12254
                     /organism="Gateway binary vector pGWB661"
                     /mol_type="other DNA"
                     /db_xref="taxon:711439"
     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    1178..1888
                     /label=tag red fluorescent protein
                     /note="tag red fluorescent protein"
     CDS             1178..1888
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    complement(1898..2022)
                     /label=attR1
                     /note="attR1"
     protein_bind    1898..2022
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2059..2089
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2143..2802
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="BAI67509.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            2143..2802
                     /gene="cat"
                     /label=cat
     CDS             3144..3449
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAI67510.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            3144..3449
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    3490..3614
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(3490..3614)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3624..4033
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      3640..3892
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3926..3942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3950..3966
                     /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(3974..4004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4019..4040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      4076..4328
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      complement(4078..4344)
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     CDS             complement(4488..5039)
                     /codon_start=1
                     /gene="bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=bar
                     /protein_id="BAI67511.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            complement(4488..5039)
                     /gene="bar"
                     /label=bar
     regulatory      complement(5067..5353)
                     /regulatory_class="promoter"
                     /note="NOS promoter"
     promoter        complement(5091..5270)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    complement(5776..5800)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     CDS             6321..7112
                     /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      7358..7946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    8132..8272
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      8616..8810
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8876..9949)
                     /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(10378..11007)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"