LOCUS       Exported                7737 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pARS3m, complete sequence.
ACCESSION   KT985053
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7737)
  AUTHORS   Thilmony R.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2015) Crop Improvement and Genetics Research Unit,
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   2  (bases 1 to 7737)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7737
                     /organism="Binary vector pARS3m"
                     /mol_type="other DNA"
                     /db_xref="taxon:1790065"
     misc_feature    1..25
                     /label=Agrobacterium right border
                     /note="Agrobacterium right border"
     misc_feature    68..235
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      236..1944
                     /regulatory_class="promoter"
                     /note="rice ubiquitin 2 promoter; RUBQ2"
     promoter        236..1944
                     /gene="RUBQ2"
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin 
                     gene RUBQ2"
     intron          983..1944
                     /note="RUBQ2 5 intron"
     CDS             1945..3204
                     /codon_start=1
                     /gene="ubiM-hptII"
                     /product="UbiM-HptII"
                     /function="confers hygromycin resistance in plants"
                     /label=ubiM-hptII
                     /note="ubiquitin monomer-hygromycin phosphotransferase II 
                     translational fusion"
                     /protein_id="AMB57378.1"
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLADYNIQKESTLHLVLRLRGGGSMKKPELTATSVEKFLIEKFDSVSDLM
                     QLSEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSE
                     SLTYCISRRSQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQY
                     TTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNN
                     VLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRL
                     RAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGC
                     VEVLADSGNRRPSTRPRAKK"
     gene            1945..3204
                     /gene="ubiM-hptII"
                     /label=ubiM-hptII
     misc_feature    1945..2170
                     /gene="ubiM-hptII"
                     /note="rice ubiquitin monomer fused to HptII is a 
                     translational enhancer that is cleaved off the mature 
                     protein in eukaryotic cells"
     CDS             2179..3204
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     regulatory      3209..3424
                     /regulatory_class="terminator"
                     /note="CaMV 35S terminator"
     polyA_signal    3223..3397
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3510..3534
                     /label=Agrobacterium left border
                     /note="Agrobacterium left border"
     rep_origin      3612..4243
                     /note="oriV; incP origin of replication"
     CDS             4455..5249
                     /codon_start=1
                     /gene="nptIII"
                     /product="NptIII"
                     /function="confers kanamycin resistance in bacteria"
                     /label=nptIII
                     /note="neomycin phosphotransferase III"
                     /protein_id="AMB57379.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            4455..5249
                     /gene="nptIII"
                     /label=nptIII
     CDS             5548..6696
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /label=trfA
                     /note="oriV trans-acting replicase"
                     /protein_id="AMB57380.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            5548..6696
                     /gene="trfA"
                     /label=trfA
     rep_origin      complement(6936..7524)
                     /direction=LEFT
                     /label=high copy-number ColE1 origin of replication
                     /note="high copy-number ColE1 origin of replication"