LOCUS       Exported               13863 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pIPKb003, complete sequence.
ACCESSION   EU161569
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13863)
  AUTHORS   Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer 
            P, Kumlehn J.
  TITLE     A set of modular binary vectors for transformation of cereals
  JOURNAL   Plant Physiol. 145 (4), 1192-1200 (2007)
  PUBMED    17981986
REFERENCE   2  (bases 1 to 13863)
  AUTHORS   Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer 
            P, Kumlehn J.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-SEP-2007) Cytogenetics and Molecular Cell Biology, 
            Leibniz Institute of Plant Genetics and Crop Plant Research, 
            Corrensstr. 3, Gatersleben D-06466, Germany
REFERENCE   3  (bases 1 to 13863)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13863
                     /organism="Binary vector pIPKb003"
                     /mol_type="other DNA"
                     /label=binary vector for over-expression in cereals
                     /note="binary vector for over-expression in cereals"
                     /db_xref="taxon:475865"
     regulatory      10..996
                     /regulatory_class="promoter"
                     /note="maize ubiquitin (Ubi-1) promoter"
     intron          997..1516
                     /note="maize ubiquitin (Ubi-1) intron 1"
     CDS             1544..2569
                     /codon_start=1
                     /gene="hpt"
                     /product="hygromycin phosphotransferase"
                     /label=hpt
                     /protein_id="ABW70076.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     gene            1544..2569
                     /gene="hpt"
                     /label=hpt
     regulatory      2594..2795
                     /regulatory_class="terminator"
                     /note="CaMV 35S terminator"
     regulatory      2828..2837
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      complement(2847..3100)
                     /regulatory_class="terminator"
                     /note="nopaline synthase terminator"
     terminator      2848..3100
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_recomb     3119..3243
                     /label=attR2
                     /note="attR2"
     protein_bind    3119..3243
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3284..3589)
                     /codon_start=1
                     /gene="ccdB"
                     /product="gyrase inhibitor"
                     /label=ccdB
                     /protein_id="ABW70074.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(3284..3589)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(3931..4590)
                     /codon_start=1
                     /gene="CmR"
                     /product="chloramphenicol resistance protein"
                     /label=CmR
                     /protein_id="ABW70075.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(3931..4590)
                     /gene="CmR"
                     /label=CmR
     promoter        complement(4644..4674)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_recomb     4699..4823
                     /label=attR1
                     /note="attR1"
     protein_bind    complement(4699..4823)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    complement(4862..6339)
                     /label=rice actin (Act-1) promoter and intron 1
                     /note="rice actin (Act-1) promoter and intron 1"
     CDS             complement(5378..5395)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(5398..6230)
                     /label=Act1 promoter
                     /note="rice actin promoter"
     misc_feature    6337..6361
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    6338..6368
                     /label=right T-DNA border
                     /note="right T-DNA border"
     rep_origin      6896..7593
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     rep_origin      7006..7594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7779..7919
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      8142..11847
                     /label=pVS1 origin of replication
                     /note="pVS1 origin of replication"
     CDS             9254..9883
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             10312..11385
                     /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      11451..11645
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             12031..13041
                     /codon_start=1
                     /gene="spec"
                     /product="streptomycin/spectinomycin adenyltransferase"
                     /label=spec
                     /protein_id="ABW70077.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            12031..13041
                     /gene="spec"
                     /label=spec
     CDS             12250..13041
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    13561..13585
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    13562..13585
                     /label=left T-DNA border
                     /note="left T-DNA border"