LOCUS       Exported                9369 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pH7WG_ASX, complete sequence.
ACCESSION   MG561372
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9369)
  AUTHORS   Lukan T, Machens F, Coll A, Baebler S, Messerschmidt K, Gruden K.
  TITLE     Plant X-tender: An extension of the AssemblX system for the assembly
            and expression of multigene constructs in plants
  JOURNAL   PLoS ONE 13 (1), e0190526 (2018)
  PUBMED    29300787
REFERENCE   2  (bases 1 to 9369)
  AUTHORS   Lukan T, Machens F, Coll A, Baebler S, Messerschmidt K, Gruden K.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-NOV-2017) Department of Biotechnology and Systems 
            Biology, National Institute of Biology, Vecna pot 111, Ljubljana 
            1000, Slovenia
REFERENCE   3  (bases 1 to 9369)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9369
                     /organism="Binary vector pH7WG_ASX"
                     /mol_type="other DNA"
                     /db_xref="taxon:2070780"
     misc_feature    join(1..115,9285..9369)
                     /label=Right T-DNA border, RB
                     /note="Right T-DNA border, RB"
     misc_feature    join(1..8454,9285..9369)
                     /label=pH7WG backbone
                     /note="pH7WG backbone"
     CDS             1331..1960
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2394..3461
                     /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"
     rep_origin      3527..3721
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    4065..4205
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4391..4979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5049..6298)
                     /label=Spectinomycin resistance cassette
                     /note="Spectinomycin resistance cassette"
     CDS             complement(5223..6155)
                     /codon_start=1
                     /gene="specR"
                     /product="SpecR"
                     /label=specR
                     /note="Spectinomycin resistance"
                     /protein_id="AUO29334.1"
                     /translation="MTCFFGVQSMPRASKQQARYAVGRCLMLWSSNDVTQQGSRPKTKL
                     NIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVR
                     LDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRND
                     ILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNS
                     PPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQ
                     EEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(5223..6155)
                     /gene="specR"
                     /label=specR
     CDS             complement(5223..6014)
                     /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"
     misc_feature    6304..6636
                     /label=left T-DNA border, LB
                     /note="left T-DNA border, LB"
     misc_feature    6542..6566
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(6640..8414)
                     /label=Hygromycin resistance cassette
                     /note="Hygromycin resistance cassette"
     terminator      6659..6911
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(7109..8131)
                     /codon_start=1
                     /gene="hygR"
                     /product="HygR"
                     /label=hygR
                     /note="Hygromycin resistance"
                     /protein_id="AUO29335.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWARTAPKSGTRARGFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     gene            complement(7109..8131)
                     /gene="hygR"
                     /label=hygR
     promoter        complement(8167..8346)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    8455..8472
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     misc_feature    8491..8540
                     /label=homology region A0
                     /note="homology region A0"
     misc_feature    8541..8546
                     /label=HindIII restriction site
                     /note="HindIII restriction site"
     misc_feature    8547..9209
                     /label=ccdB cassette
                     /note="ccdB cassette"
     CDS             8876..9181
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    9210..9215
                     /label=HindIII restriction site
                     /note="HindIII restriction site"
     misc_feature    9216..9265
                     /label=homology region B0
                     /note="homology region B0"
     misc_feature    complement(9266..9283)
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     misc_feature    9346..1
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"