LOCUS       Exported               10818 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUB-DEST, complete sequence.
ACCESSION   KC441463
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10818)
  AUTHORS   Grefen C, Donald N, Hashimoto K, Kudla J, Schumacher K, Blatt MR.
  TITLE     A ubiquitin-10 promoter-based vector set for fluorescent protein 
            tagging facilitates temporal stability and native protein 
            distribution in transient and stable expression studies
  JOURNAL   Plant J. 64 (2), 355-365 (2010)
  PUBMED    20735773
REFERENCE   2  (bases 1 to 10818)
  AUTHORS   Grefen C, Donald N, Hashimoto K, Kudla J, Schumacher K, Blatt MR.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-NOV-2012) Plant Sciences Group, University of Glasgow,
            University Ave, Glasgow G12 8QQ, United Kingdom
REFERENCE   3  (bases 1 to 10818)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10818
                     /organism="Cloning vector pUB-DEST"
                     /mol_type="other DNA"
                     /db_xref="taxon:1298537"
     regulatory      1..639
                     /regulatory_class="promoter"
                     /note="ubiquitin 10 promoter from Arabidopsis thaliana 
                     Columbia-6 ecotype (At4g05050)"
     misc_recomb     668..792
                     /label=attR1 gateway recombination site
                     /note="attR1 gateway recombination site"
     protein_bind    668..792
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        817..847
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             901..1560
                     /codon_start=1
                     /product="chloramphenicol acetyl transferase"
                     /label=chloramphenicol acetyl transferase
                     /note="CmR; confers resistance to chloramphenicol"
                     /protein_id="AGG35610.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1902..2207
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /protein_id="AGG35611.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_recomb     complement(2248..2372)
                     /label=attR2 gateway recombination site
                     /note="attR2 gateway recombination site"
     protein_bind    complement(2248..2372)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2449..2657
                     /regulatory_class="terminator"
                     /note="T35S terminator"
     promoter        2794..2973
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             3015..3566
                     /codon_start=1
                     /product="phosphinothricin N-acetyltransferase"
                     /label=phosphinothricin N-acetyltransferase
                     /note="confers Basta herbicide resistance"
                     /protein_id="AGG35612.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      3586..3838
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3861..4193
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    3931..3955
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             4257..5267
                     /codon_start=1
                     /product="aminoglycoside resistance protein"
                     /label=aminoglycoside resistance protein
                     /note="AadA; confers resistance to spectinomycin"
                     /protein_id="AGG35613.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             4476..5267
                     /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      5514..6102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5718..6398
                     /note="pBR322; replication in Escherichia coli"
     misc_feature    6288..6428
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      6772..6966
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      6808..9401
                     /note="pVS1; replication in Agrobacterium tumefaciens"
     CDS             complement(7032..8099)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     CDS             complement(8533..9162)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    10378..10526
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    10492..10516
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    10681..10702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10717..10747
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10755..10771
                     /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)."
     primer_bind     10779..10795
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"