LOCUS       Exported                6675 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pKDH, complete sequence.
ACCESSION   KF041008
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6675)
  AUTHORS   Sahoo DK, Maiti IB.
  TITLE     Complete sequence of binary plant expression vector pKDH with 
            Hygromycin selectable marker gene
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6675)
  AUTHORS   Sahoo DK, Maiti IB.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-MAY-2013) Plant Genetic Engineering Research and 
            Services, Kentucky Tobacco Research & Development Center, College of
            Agriculture, University of Kentucky, 1401 University Drives, 
            Lexington, KY 40546-0236, USA
REFERENCE   3  (bases 1 to 6675)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6675
                     /organism="Binary vector pKDH"
                     /mol_type="other DNA"
                     /country="USA"
                     /db_xref="taxon:1389449"
     misc_feature    complement(13..169)
                     /note="left T-DNA border of Ti-plasmid; similar to INSD 
                     accession AF139061"
     misc_feature    66..90
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    170..228
                     /label=1st polylinker/multiple cloning site
                     /note="1st polylinker/multiple cloning site"
     misc_feature    229..287
                     /label=2nd polylinker/multiple cloning site
                     /note="2nd polylinker/multiple cloning site"
     terminator      287..928
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     3UTR           288..927
                     /gene="rbcSE9"
                     /label=similar to INSD accession X00806
                     /note="similar to INSD accession X00806"
     gene            288..927
                     /gene="rbcSE9"
                     /label=rbcSE9
                     /note="derived from pea"
     gene            934..2495
                     /gene="nopaline synthase"
                     /label=nopaline synthase
     regulatory      934..1217
                     /regulatory_class="promoter"
                     /gene="nopaline synthase"
                     /note="similar to INSD accession V00087"
     promoter        1017..1196
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             1218..2246
                     /codon_start=1
                     /gene="nopaline synthase"
                     /product="hygromycin resistance protein"
                     /label=nopaline synthase
                     /note="HygR; codon-optimized with plant bias codons"
                     /protein_id="AGU26639.1"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KK"
     3UTR           2247..2495
                     /gene="nopaline synthase"
                     /label=similar to INSD accession V00087
                     /note="similar to INSD accession V00087"
     misc_feature    2497..2550
                     /label=3rd linker/multiple cloning site
                     /note="3rd linker/multiple cloning site"
     misc_feature    complement(2551..2712)
                     /note="right T-DNA border of Ti-plasmid; similar to INSD 
                     accession J01826"
     misc_feature    2648..2672
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2731..3398)
                     /direction=LEFT
                     /note="ColE1 origin of replication; similar to XM2327"
     rep_origin      complement(2787..3375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     3UTR           complement(3399..3542)
                     /label=similar to XM2327
                     /note="similar to XM2327"
     gene            complement(3542..4534)
                     /gene="AmpR"
                     /label=AmpR
                     /note="bla"
     CDS             complement(3546..4406)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin; similar to XM2327"
                     /protein_id="AGU26640.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      complement(4407..4534)
                     /regulatory_class="promoter"
                     /gene="AmpR"
                     /note="similar to XM2327"
     promoter        complement(4407..4511)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4541..5164
                     /note="pRK2-OriV; similar to INSD accession AF139061"
     rep_origin      4639..5170
                     /label=oriV
                     /note="incP origin of replication"
     gene            5171..6675
                     /gene="trfA"
                     /label=trfA
                     /note="plasmid pRK2trfA region; transacting replication"
     regulatory      5171..5474
                     /regulatory_class="promoter"
                     /gene="trfA"
                     /note="similar to INSD accession X00713"
     CDS             5475..6623
                     /codon_start=1
                     /gene="trfA"
                     /product="trfA"
                     /label=trfA
                     /note="codon-optimized for expression in bacteria; similar 
                     to INSD accession X00713"
                     /protein_id="AGU26641.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     3UTR           6624..6675
                     /gene="trfA"
                     /label=similar to INSD accession X00713
                     /note="similar to INSD accession X00713"