LOCUS       Exported               12117 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPIPRA540, complete sequence.
ACCESSION   KP638477
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12117)
  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  JOURNAL   Unpublished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 12117)
  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 12117)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12117
                     /organism="Cloning vector pPIPRA540"
                     /mol_type="other DNA"
                     /db_xref="taxon:1642675"
     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="AKE42810.1"
                     /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      1869..2812
                     /regulatory_class="promoter"
                     /note="FMV34S"
     misc_feature    2868..4921
                     /label=partial GUS coding region
                     /note="partial GUS coding region"
     CDS             join(2869..2883,3074..4891)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=GUS
                     /note="This version of the gusA gene has a 5 extension 
                     with a catalase intron to ensure expression in plants but 
                     not bacteria."
                     /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES
                     ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK
                     VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK
                     KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV
                     ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELEVTAKSQTESDIYPLRVGIRS
                     VAVKGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP
                     YAEEMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA
                     IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD
                     AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT
                     LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK
                     GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     intron          2884..3073
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             4898..4915
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      4946..5246
                     /regulatory_class="terminator"
                     /note="pea E9"
     misc_feature    5325..5349
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    5462..5569
                     /label=T-DNA right border
                     /note="T-DNA right border"
     rep_origin      5928..9380
                     /label=Agrobacterium pVS1
                     /note="Agrobacterium pVS1"
     CDS             6801..7430
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7859..8932
                     /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      8998..9192
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      9525..10655
                     /label=E. coli ColE1
                     /note="E. coli ColE1"
     misc_feature    9666..9806
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9992..10580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(11045..11839)
                     /codon_start=1
                     /gene="aadA"
                     /product="AadA"
                     /label=aadA
                     /note="confers kanamycin resistance"
                     /protein_id="AKE42811.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(11045..11839)
                     /gene="aadA"
                     /label=aadA