LOCUS       Exported               13015 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPIPRA719, complete sequence.
ACCESSION   KP638479
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13015)
  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  JOURNAL   Unpublished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 13015)
  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 13015)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13015
                     /organism="Cloning vector pPIPRA719"
                     /mol_type="other DNA"
                     /db_xref="taxon:1642677"
     regulatory      16..419
                     /regulatory_class="promoter"
                     /note="mas"
     promoter        26..406
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             420..1142
                     /codon_start=1
                     /gene="ipt"
                     /product="isopentenyl phosphotransferase"
                     /label=ipt
                     /note="involved in condensation of AMP and 
                     isopentenylpyrophosphate to form isopentenyl-AMP"
                     /protein_id="AKE42814.1"
                     /translation="MDLHLIFGPTCTGKTTTAIALAQQTGLPVLSLDRVQCCPQLSTGS
                     GRPTVEELKGTTRLYLDDRPLVEGIIAAKQAHHRLIEEVYNHEANGGLILEGGSTSLLN
                     CMARNSYWSADFRWHIIRHKLPDQETFMKAAKARVKQMLHPAAGHSIIQELVYLWNEPR
                     LRPILKEIDGYRYAMLFASQNQITADMLLQLDANMEGKLINGIAQEYFIHARQQEQKFP
                     QVNAAAFDGFEGHPFGMY"
     gene            420..1142
                     /gene="ipt"
                     /label=ipt
     regulatory      1494..1770
                     /regulatory_class="terminator"
                     /note="mas"
     terminator      1505..1757
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    1782..1949
                     /label=T-DNA left border
                     /note="T-DNA left border"
     regulatory      1974..2372
                     /regulatory_class="promoter"
                     /note="mas"
     promoter        1984..2364
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             2378..3175
                     /codon_start=1
                     /gene="NPTII"
                     /product="neomycin phosphotransferase"
                     /label=NPTII
                     /note="confers kanamycin resistance"
                     /protein_id="AKE42815.1"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            2378..3175
                     /gene="NPTII"
                     /label=NPTII
     regulatory      3182..3458
                     /regulatory_class="terminator"
                     /note="mas"
     terminator      3193..3445
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     regulatory      3502..4445
                     /regulatory_class="promoter"
                     /note="FMV34S"
     CDS             4512..5795
                     /codon_start=1
                     /gene="codA"
                     /product="cytosine deaminase"
                     /label=codA
                     /note="negative selection marker"
                     /protein_id="AKE42816.1"
                     /translation="MSNNALQTIINARLPGEEGLWQIHLQDGKISAIDAQSGVMPITEN
                     SLDAEQGLVIPPFVEPHIHLDTTQTAGQPNWNQSGTLFEGIERWAERKALLTHDDVKQR
                     AWQTLKWQIANGIQHVRTHVDVSDATLTALKAMLEVKQEVAPWIDLQIVAFPQEGILSY
                     PNGEALLEEALRLGADVVGAIPHFEFTREYGVESLHKTFALAQKYDRLIDVHCDEIDDE
                     QSRFVETVAALAHHEGMGARVTASHTTAMHSYNGAYTSRLFRLLKMSGINFVANPLVNI
                     HLQGRFDTYPKRRGITRVKEMLESGINVCFGHDDVFDPWYPLGTANMLQVLHMGLHVCQ
                     LMGYGQINDGLNLITHHSARTLNLQDYGIAAGNSANLIILPAENGFDALRRQVPVRYSV
                     RGGKVIASTQPAQTTVYLEQPEAIDYKR"
     gene            4512..5795
                     /gene="codA"
                     /label=codA
     regulatory      5810..6110
                     /regulatory_class="terminator"
                     /note="pea E9"
     misc_feature    6154..6362
                     /label=T-DNA left border
                     /note="T-DNA left border"
     rep_origin      6528..10280
                     /label=Agrobacterium pVS1
                     /note="Agrobacterium pVS1"
     CDS             7699..8328
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8757..9830
                     /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      9896..10090
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      10425..11555
                     /label=E. coli ColE1
                     /note="E. coli ColE1"
     misc_feature    10564..10704
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(10890..11478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(11943..12737)
                     /codon_start=1
                     /gene="aadA"
                     /product="AadA"
                     /label=aadA
                     /note="confers kanamycin resistance"
                     /protein_id="AKE42817.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(11943..12737)
                     /gene="aadA"
                     /label=aadA