LOCUS       Exported               10559 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPIPRA728, complete sequence.
ACCESSION   KP638480
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10559)
  AUTHORS   Figueroa-Balderas RE, Chi-Ham CL, Le H, Tricoli D, Sandman J, 
            Johnson D, Reich J, Bennett AB.
  TITLE     Constitutive expression of eIF5A3 increases high quality biomass 
            yield in an elite alfalfa cultivar
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10559)
  AUTHORS   Figueroa-Balderas RE, Chi-Ham CL, Le H, Tricoli D, Sandman J, 
            Johnson D, Reich J, Bennett AB.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2015) Plant Sciences, UC Davis, One shields 
            Avenue, Davis, CA 95616, USA
REFERENCE   3  (bases 1 to 10559)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10559
                     /organism="Cloning vector pPIPRA728"
                     /mol_type="other DNA"
                     /db_xref="taxon:1642678"
     misc_feature    13..337
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    245..269
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      348..755
                     /regulatory_class="promoter"
                     /note="mas"
     promoter        362..742
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             756..1553
                     /codon_start=1
                     /gene="NPTII"
                     /product="neomycin phosphotransferase"
                     /label=NPTII
                     /note="confers kanamycin resistance"
                     /protein_id="AKE42818.1"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            756..1553
                     /gene="NPTII"
                     /label=NPTII
     regulatory      1560..1836
                     /regulatory_class="terminator"
                     /note="mas"
     terminator      1571..1823
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     regulatory      1880..2823
                     /regulatory_class="promoter"
                     /note="FMV34S"
     CDS             2875..3354
                     /codon_start=1
                     /gene="eIF5A3"
                     /product="eukaryotic translation initiation factor 5A"
                     /label=eIF5A3
                     /note="eIF5A3; derived from Populus deltoides"
                     /protein_id="AKE42819.1"
                     /translation="MSDEEQHFESKADAGASKTYPQQAGTIRKSGYIVIKNRPCKVVEV
                     STSKTGKHGHAKCHFVAIDIFNGKKLEDIVPSSHNCDVPHVTRTDYQLIDISEDGFVSL
                     LTENGNTKDDLRLPTDESLLSQIKDGFGEGKDLVVTVMSSMGEEQICALKDVGPK"
     gene            2875..3354
                     /gene="eIF5A3"
                     /label=eIF5A3
                     /note="PdeIF5A3"
     regulatory      3377..3680
                     /regulatory_class="terminator"
                     /note="pea E9"
     misc_feature    3711..3904
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    3767..3791
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4070..7822
                     /label=Agrobacterium pVS1
                     /note="Agrobacterium pVS1"
     CDS             5243..5872
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6301..7374
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      7440..7634
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      7967..9097
                     /label=E. coli ColE1
                     /note="E. coli ColE1"
     misc_feature    8108..8248
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(8434..9022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9487..10281)
                     /codon_start=1
                     /gene="aadA"
                     /product="AadA"
                     /label=aadA
                     /note="confers kanamycin resistance"
                     /protein_id="AKE42820.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(9487..10281)
                     /gene="aadA"
                     /label=aadA