LOCUS       Exported                7081 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pSiM24, complete sequence.
ACCESSION   KF032933
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7081)
  AUTHORS   Sahoo DK, Dey N, Maiti IB.
  TITLE     pSiM24 Is a Novel Versatile Gene Expression Vector for Transient 
            Assays As Well As Stable Expression of Foreign Genes in Plants
  JOURNAL   PLoS ONE 9 (6), E98988 (2014)
  PUBMED    24897541
REFERENCE   2  (bases 1 to 7081)
  AUTHORS   Maiti IB, Sahoo DK.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAY-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 7081)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7081
                     /organism="Binary vector pSiM24"
                     /mol_type="other DNA"
                     /country="USA"
                     /db_xref="taxon:1392513"
     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..234
                     /label=1st polylinker/multiple cloning site
                     /note="1st polylinker/multiple cloning site"
     regulatory      235..860
                     /regulatory_class="promoter"
                     /note="M24 promoter; derived from Mirabilis Mosaic Virus 
                     (MMV)"
     misc_feature    861..919
                     /label=2nd polylinker/multiple cloning site
                     /note="2nd polylinker/multiple cloning site"
     terminator      919..1560
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     regulatory      920..1559
                     /regulatory_class="polyA_signal_sequence"
                     /note="rbcSE9 terminator sequence; derived from pea"
     regulatory      complement(1572..1820)
                     /regulatory_class="terminator"
                     /note="nopaline synthase terminator; NosT; Nos-terminator"
     gene            complement(1821..2902)
                     /gene="nos-nptII"
                     /label=nos-nptII
     CDS             complement(1821..2618)
                     /codon_start=1
                     /gene="nos-nptII"
                     /product="kanamycin resistance protein"
                     /label=nos-nptII
                     /note="neomycin phosphotransferase II; KanR; 
                     codon-optimized for expression in plants; similar to INSD 
                     accession V00618"
                     /protein_id="AGU26662.1"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(2619..2902)
                     /regulatory_class="promoter"
                     /gene="nos-nptII"
                     /note="Nos promoter; similar to INSD accession XV00087"
     promoter        complement(2640..2819)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    2903..2956
                     /label=3rd linker/multiple cloning site
                     /note="3rd linker/multiple cloning site"
     misc_feature    complement(2957..3118)
                     /note="right T-DNA border; similar to INSD accession 
                     J01826"
     misc_feature    3054..3078
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3137..3804)
                     /direction=LEFT
                     /note="ColE1 origin of replication; similar to INSD 
                     accession XM2327"
     rep_origin      complement(3193..3781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(3805..4940)
                     /gene="bla"
                     /label=bla
                     /note="ampR"
     3UTR           complement(3805..3951)
                     /gene="bla"
                     /label=similar to INSD accession XM2327
                     /note="similar to INSD accession XM2327"
     CDS             complement(3952..4812)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="confers resistance to ampicillin; similar to INSD 
                     accession XM2327"
                     /protein_id="AGU26661.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      complement(4813..4940)
                     /regulatory_class="promoter"
                     /gene="bla"
                     /note="similar to INSD accession XM2327"
     promoter        complement(4813..4917)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4947..5570
                     /note="pRK2-Ori V; similar to INSD accession AF139061"
     rep_origin      5045..5576
                     /label=oriV
                     /note="incP origin of replication"
     gene            5577..7081
                     /gene="pRK2-trfA"
                     /label=pRK2-trfA
     regulatory      5577..5880
                     /regulatory_class="promoter"
                     /gene="pRK2-trfA"
     CDS             5881..7029
                     /codon_start=1
                     /gene="pRK2-trfA"
                     /product="pRK2-trfA"
                     /label=pRK2-trfA
                     /note="trans-acting replication; similar to INSD accession 
                     X00713"
                     /protein_id="AGU26663.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     3UTR           7030..7081
                     /gene="pRK2-trfA"
                     /label=trfA-terminator
                     /note="trfA-terminator"