LOCUS       Exported                9753 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCAMBIA_ASX, complete sequence.
ACCESSION   MG561370
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9753)
  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 9753)
  AUTHORS   Lukan T, Machens F, Coll A, Baebler S, Messerschmidt K, Gruden K.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-NOV-2017) Department of Biotechnology and Systems 
            Biology, National Institute of Biology, Vecna pot 111, Ljubljana 
            1000, Slovenia
REFERENCE   3  (bases 1 to 9753)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9753
                     /organism="Binary vector pCAMBIA_ASX"
                     /mol_type="other DNA"
                     /db_xref="taxon:2070779"
     misc_feature    join(15..20,684..689)
                     /label=HindIII restriction site
                     /note="HindIII restriction site"
     misc_feature    21..683
                     /label=ccdB cassette
                     /note="ccdB cassette"
     CDS             350..655
                     /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    690..739
                     /label=homology region B0
                     /note="homology region B0"
     misc_feature    complement(740..757)
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     misc_feature    759..9681
                     /note="pCAMBIA1300 backbone, derived from INSD accession 
                     AF234296.1"
     primer_bind     complement(762..778)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    981..1005
                     /note="RB T-DNA repeat; right border repeat from nopaline 
                     C58 T-DNA"
     CDS             2305..2934
                     /codon_start=1
                     /product="stability protein"
                     /label=stability protein
                     /note="derived from Pseudomonas plasmid pVS1; pVS1 StaA"
                     /protein_id="AUO29329.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    3363..4436
                     /product="replication protein from Pseudomonas plasmid 
                     pVS1, pVS1 RepA"
     CDS             3363..4436
                     /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"
     rep_origin      4502..4696
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000), pVS1 oriV"
     misc_feature    5040..5180
                     /label=basis of mobility region from pBR322
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5366..5954)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6041..6835)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="confers resistance to kanamycin"
                     /protein_id="AUO29330.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    7260..7284
                     /note="LB T-DNA repeat, left border repeat from nopaline 
                     C58 T-DNA"
     misc_feature    7362..7536
                     /note="CaMV poly(A) signal, cauliflower mosaic virus 
                     polyadenylation signal"
     polyA_signal    7362..7536
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(7576..8601)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=hygromycin B phosphotransferase
                     /note="confers resistance to hygromycin"
                     /protein_id="AUO29331.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     regulatory      complement(8669..9346)
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter, cauliflower mosaic virus 35S 
                     promoter with a duplicated enhancer region"
     promoter        complement(8669..9346)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    9537..9558
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9573..9603
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9611..9627
                     /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     9635..9651
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9682..9699
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     misc_feature    9718..14
                     /label=homology region A0
                     /note="homology region A0"