LOCUS       Exported               12945 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pKM24KH, complete sequence.
ACCESSION   HM036220
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12945)
  AUTHORS   Maiti IB, Sahoo DK, Raha S, Pattanaik S.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-MAR-2010) KTRDC Plant Genetic Engineering Research & 
            Services, University of Kentucky, 1401 University Drive, Lexington, 
            KY 40546, USA
REFERENCE   2  (bases 1 to 12945)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12945
                     /organism="Binary vector pKM24KH"
                     /mol_type="other DNA"
                     /db_xref="taxon:762950"
     gene            complement(1..2000)
                     /gene="nos"
                     /gene_synonym="nptII"
                     /label=nos
     3UTR           complement(1..879)
                     /gene="nos"
                     /gene_synonym="nptII"
     regulatory      complement(1..243)
                     /regulatory_class="terminator"
                     /gene="nos"
                     /gene_synonym="nptII"
     CDS             complement(880..1701)
                     /codon_start=1
                     /gene="nos"
                     /gene_synonym="nptII"
                     /product="neomycin phosphotransferase II"
                     /label=nos
                     /note="kanamycin resistance protein; nos-nptII"
                     /protein_id="ADG63448.1"
                     /translation="MAITLSATSLPISARIRAGSPAAWVERLFGYDWAQQTIGCSDAAV
                     FRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLG
                     EVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVD
                     QDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLG
                     VADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(1702..2000)
                     /regulatory_class="promoter"
                     /gene="nos"
                     /gene_synonym="nptII"
     promoter        complement(1702..1885)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(1738..1917)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    2001..2162
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    2041..2065
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     repeat_region   2060..2085
                     /label=right T-DNA border repeat
                     /note="right T-DNA border repeat"
     rep_origin      2553..3184
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      2555..3184
                     /note="OriV; incP"
     misc_feature    3946..6392
                     /note="teracyline resistance (tetA-terR) region; similar to
                     GenBank Accession Number X75761"
     CDS             complement(4248..4898)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance protein"
                     /label=tetR
                     /note="TetR; Tetracycline resistance regulation"
                     /protein_id="ADG63449.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     gene            complement(4248..4898)
                     /gene="tetR"
                     /label=tetR
     CDS             5004..6203
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline resistance protein"
                     /label=tetA
                     /note="TetA"
                     /protein_id="ADG63450.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            5004..6203
                     /gene="tetA"
                     /label=tetA
     CDS             complement(7476..8624)
                     /codon_start=1
                     /gene="trfA"
                     /product="trans-acting replication protein"
                     /label=trfA
                     /note="TrfA; trans-acting replication"
                     /protein_id="ADG63451.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(7476..8624)
                     /gene="trfA"
                     /label=trfA
     CDS             complement(8896..9267)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /note="TraJ"
                     /protein_id="ADG63452.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(8896..9267)
                     /gene="traJ"
                     /label=traJ
     oriT            complement(9300..9409)
                     /direction=LEFT
                     /note="oriT; incP origin of transfer"
     misc_feature    9891..10040
                     /label=left T-DNA border of Ti-plasmid
                     /note="left T-DNA border of Ti-plasmid"
     misc_feature    9964..9988
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     repeat_region   10041..10675
                     /label=left T-DNA border repeat
                     /note="left T-DNA border repeat"
     regulatory      10041..10675
                     /regulatory_class="promoter"
                     /note="M24 promoter from Mirabilis Mosaic Virus (MMV)"
     misc_feature    10676..10720
                     /label=polylinker
                     /note="polylinker"
     terminator      10720..11361
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     regulatory      10721..11366
                     /regulatory_class="terminator"
                     /note="terminator; 3UTR sequence of rbcSE9"
     regulatory      complement(11367..11656)
                     /regulatory_class="promoter"
                     /note="nos"
     promoter        11450..11629
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             11657..12682
                     /codon_start=1
                     /gene="hph"
                     /product="hygromycin phosphotransferase"
                     /label=hph
                     /note="Hph; hygromycin resistance protein"
                     /protein_id="ADG63453.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDQMQLSEGEESRAFSFDVRGRG
                     YVLRVNSCADGSYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     G"
     gene            11657..12682
                     /gene="hph"
                     /label=hph
     regulatory      12683..12945
                     /regulatory_class="terminator"
                     /note="terminator; 3UTR of the nos gene"