LOCUS       Exported                9170 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pC1300intB, complete sequence.
ACCESSION   AF294977
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9170)
  AUTHORS   Ouwerkerk PBF.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-AUG-2000) Dept. of Molecular Cell Biology, Institute 
            for Molecular Plant Sciences, Wassenaarseweg 64, Leiden 2333 AL, 
            Netherlands
REFERENCE   2  (bases 1 to 9170)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9170
                     /organism="Cloning vector pC1300intB"
                     /mol_type="genomic DNA"
                     /note="derived from Binary vector pCAMBIA-1300 presented in
                     GenBank Accession Number AF234296"
                     /db_xref="taxon:142850"
     misc_feature    1..9170
                     /note="presense of the catalase-1 intron in the hptII gene 
                     confers higher resistance towards hygromycin in transgenic 
                     rice and allows an easier killing of Agrobacterium 
                     tumefaciens during regeneration of transgenic plants; 
                     alternative for pC1300intB is pC1300intA (Genbank Accession
                     Number AF294976) or pC1300intC (Genbank Accession Number 
                     AF294978); cloning source for the Cat-1 intron was binary 
                     vector pWBVec8 (Wang et al., 1998, Acta Horticultur. 
                     461:401-407)"
     misc_feature    1..57
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(61..77)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    280..304
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    299..324
                     /label=right border repeat from C58 T-DNA
                     /note="right border repeat from C58 T-DNA"
     CDS             1604..2233
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2667..3734
                     /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      3800..3994
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    4338..4478
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4664..5252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5339..6133)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6558..6583
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    6558..6582
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     3UTR           complement(6650..6869)
                     /label=CaMV (polyA signal)
                     /note="CaMV (polyA signal)"
     polyA_signal    6660..6834
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(join(6885..7577,7768..8100))
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /label=hptII
                     /note="hygromycin resistance"
                     /protein_id="AAG38026.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(6885..8100)
                     /gene="hptII"
                     /label=hptII
     exon            complement(6885..7577)
                     /gene="hptII"
                     /number=2
     intron          7572..7761
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     intron          complement(7578..7767)
                     /gene="hptII"
                     /number=1
                     /note="derived from Ricinus communis catalase-1 intron 
                     presented in Genbank Accession Number D21161"
     exon            complement(7768..8100)
                     /gene="hptII"
                     /number=1
     regulatory      complement(8147..8927)
                     /regulatory_class="promoter"
                     /note="CaMV35S; 35S promoter from CaMV"
     promoter        complement(8179..8856)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    9047..9068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9083..9113
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9121..9137
                     /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     9145..9161
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             9157..220
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"