LOCUS       Exported               14268 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pKM24KHibm10, complete sequence.
ACCESSION   KF951258
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14268)
  AUTHORS   Sahoo DK, Raha S, Hall JT, Maiti IB.
  TITLE     Overexpression of the synthetic chimeric native-T-phylloplanin-GFP 
            genes optimized for monocot and dicot plants renders enhanced 
            resistance to blue mold disease in tobacco (N. tabacum L.)
  JOURNAL   ScientificWorldJournal 2014, 601314 (2014)
  PUBMED    24778589
REFERENCE   2  (bases 1 to 14268)
  AUTHORS   Maiti IB, Sahoo DK, Raha S.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-DEC-2013) KTRDC, Plant Genetic Engineering Research 
            and Services, College of Agriculture, University of Kentucky, 1401 
            University Drive, Lexington, KY 40546, USA
REFERENCE   3  (bases 1 to 14268)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..14268
                     /organism="Binary vector pKM24KHibm10"
                     /mol_type="other DNA"
                     /country="USA"
                     /note="plant gene expression vector with a fused 
                     antifungal-GFP gene"
                     /db_xref="taxon:1468452"
     regulatory      1..641
                     /regulatory_class="promoter"
                     /note="M24 promoter from Mirabilis Mosaic Virus (MMV), Ref:
                     US Patent No. 64205470; Plant Mol. Biol. 40: 771-782 
                     (1999); Transgenics 3: 61-70 (1999)"
     regulatory      642..691
                     /regulatory_class="enhancer"
                     /note="5-untranslated region of Alfalfa mosaic virus 
                     (AlMV) RNA 4. ref: Nature 325: 622-625(1987)"
     CDS             692..1981
                     /codon_start=1
                     /gene="fused T-phylloplanin-GFP"
                     /product="fused T-phylloplanin-GFP"
                     /label=fused T-phylloplanin-GFP
                     /note="antifungal protein; chimeric synthetic fused gene 
                     T-phylloplanin-GFP, optimized for monocot (brent grass) 
                     plant bias codons; resistant to resistance to blue mold, a 
                     plant pathogen"
                     /protein_id="AHK23745.1"
                     /translation="MGANKLFLVCATFALCFLLTNAPMGASAKIFLIFLLAALIATPAA
                     FAILVPTLVSTHISGLVFCSVNGNLDVINGLSPQVFPNASVQLRCGATNVISSTITNGS
                     GAFSLAVNTFPLLNCNLVVATPLSTCNATLQSVGRLASSLRLVNITLGSGTGLIRVGLA
                     PTGFILNLNINAIGPMYNQYQYPNGSTMGSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALS
                     KDPNEKRDHMVLLEFVTAAGITHGMDELYK"
     gene            692..1981
                     /gene="fused T-phylloplanin-GFP"
                     /label=fused T-phylloplanin-GFP
     sig_peptide     692..760
                     /gene="fused T-phylloplanin-GFP"
                     /note="apoplast/cell wall/plasma membrane targeting protein
                     sequence (aTPS) of Arabidopsis 2S2 protein gene. Ref: Plant
                     Physiol. 155: 2036-2048 (2011); Proc Natl Acad Sci, USA, 
                     104: 1430-1435 (2007)"
     misc_feature    761..1213
                     /gene="fused T-phylloplanin-GFP"
                     /note="synthetic T-phylloplanin sequence, (GenBank 
                     accession No. AY705384), full-length with its own signal 
                     peptide, is optimized for monocot plant (brent grass) bias 
                     codons"
     misc_feature    1214..1261
                     /gene="fused T-phylloplanin-GFP"
                     /note="linker sequence used to fuse T-Phylloplanin and GFP 
                     genes in frame"
     misc_feature    1262..1981
                     /gene="fused T-phylloplanin-GFP"
                     /note="synthetic GFP gene optimized for monocot plant bias 
                     codons"
     regulatory      1982..2649
                     /regulatory_class="terminator"
                     /note="pea rbcSE9 terminator sequence"
     terminator      2003..2644
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    2650..14268
                     /label=similar to GenBank Accession No. HM036220
                     /note="similar to GenBank Accession No. HM036220"
     promoter        2733..2912
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             2940..3965
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDQMQLSEGEESRAFSFDVRGRG
                     YVLRVNSCADGSYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     G"
     CDS             complement(5108..5929)
                     /codon_start=1
                     /gene="nos-nptII"
                     /product="fusion between an N-terminal peptide of nopaline 
                     synthase and Tn5 aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAITLSATSLPISARIRAGSPAAWVERLFGYDWAQQTIGCSDAAV
                     FRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLG
                     EVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVD
                     QDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLG
                     VADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5930..6113)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5966..6145)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6269..6293
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      6781..7412
                     /label=oriV
                     /note="incP origin of replication"
     CDS             complement(8476..9126)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             9232..10431
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(11704..12852)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(13124..13495)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            13528..13637
                     /note="incP origin of transfer"
     misc_feature    14192..14216
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"