LOCUS       Exported                9937 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pC1300intB-35SnosEX, complete sequence.
ACCESSION   AY560325
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9937)
  AUTHORS   Ouwerkerk PBF., Meijer AH.
  TITLE     Binary expression vector for use in plants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9937)
  AUTHORS   Ouwerkerk PBF., Meijer AH.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2004) Molecular Cell Biology, Institute of 
            Biology, Leiden University, Wassenaarseweg 64, Leiden 2333 AL, The 
            Netherlands
REFERENCE   3  (bases 1 to 9937)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9937
                     /organism="Cloning vector pC1300intB-35SnosEX"
                     /mol_type="other DNA"
                     /note="derived from pC1300intB (Genbank Accession Number 
                     AF294977)"
                     /db_xref="taxon:267430"
     terminator      52..304
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      complement(52..303)
                     /regulatory_class="terminator"
                     /note="3NOS; terminator signal of nopaline synthase gene"
     misc_feature    complement(322..347)
                     /note="multiple cloning site; SpeI, XhoI, EcoRI, SalI are 
                     unique"
     misc_feature    349..404
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     regulatory      complement(425..789)
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     promoter        complement(426..472)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(828..844)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1047..1071
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    1066..1091
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"
     misc_feature    complement(2132..3132)
                     /label=STA region from pVS1 plasmid
                     /note="STA region from pVS1 plasmid"
     CDS             2371..3000
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3434..4501
                     /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      complement(3725..4725)
                     /direction=LEFT
                     /note="pVS1-REP; replication origin from pVS1"
     rep_origin      4567..4761
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    5105..5245
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(5135..5395)
                     /label=bom site from pBR322
                     /note="bom site from pBR322"
     rep_origin      complement(5431..6019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(5535..5815)
                     /direction=LEFT
                     /label=pBR322 origin of replication
                     /note="pBR322 origin of replication"
     CDS             complement(6106..6900)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside transferase"
                     /label=aadA
                     /note="AadA; kanamycin resistance gene from pIG121Hm"
                     /protein_id="AAS73283.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(6106..6900)
                     /gene="aadA"
                     /label=aadA
     misc_feature    7325..7350
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    7325..7349
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      complement(7417..7936)
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     3UTR           complement(7417..7636)
                     /label=CaMV polyA signal
                     /note="CaMV polyA signal"
     polyA_signal    7427..7601
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(join(7652..8338,8529..8867))
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /label=hptII
                     /note="HptII"
                     /protein_id="AAS73284.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(7652..8867)
                     /gene="hptII"
                     /label=hptII
     exon            complement(7652..8338)
                     /gene="hptII"
                     /number=2
     intron          complement(8339..8534)
                     /gene="hptII"
                     /note="derived from Ricinus communis catalase-1 intron 
                     presented in Genbank Accession Number D21161; confers 
                     higher resistance towards hygromycin in transgenic rice and
                     allows easier killing of Agrobacterium tumefaciens during 
                     regeneration of transgenic plants"
     intron          8339..8528
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     exon            complement(8529..8867)
                     /gene="hptII"
                     /number=1
     promoter        complement(8946..9623)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    9814..9835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9850..9880
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9888..9904
                     /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     9912..9928
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"