LOCUS       Exported                9773 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAGW570, complete sequence.
ACCESSION   MF401557
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9773)
  AUTHORS   Biswal AK, Atmodjo MA, Pattathil S, Amos RA, Yang X, Winkeler K, 
            Collins C, Mohanty SS, Ryno D, Tan L, Gelineo-Albersheim I, Hunt K, 
            Sykes RW, Turner GB, Ziebell A, Davis MF, Decker SR, Hahn MG, Mohnen
            D.
  TITLE     Increased xylan production by overexpression of 
            GalactUronosylTransferase12 (GAUT12) results in increased 
            recalcitrance and decreased growth in Populus woody biofuel 
            feedstock
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9773)
  AUTHORS   Rottmann W, Winkeler K, Biswal AK, Yang X.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-JUN-2017) Biochemistry and Molecular Biology; Complex 
            Carbohydrate Research Center, The University of Georgia, 315 
            Riverbend Rd, Athens, GA 30602, USA
REFERENCE   3  (bases 1 to 9773)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9773
                     /organism="Expression vector pAGW570"
                     /mol_type="other DNA"
                     /db_xref="taxon:2060070"
     CDS             907..1701
                     /codon_start=1
                     /product="NptIII"
                     /label=NptIII
                     /note="kanamycin resistance"
                     /protein_id="AUG68173.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             2000..3148
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    3505..3529
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    3506..3529
                     /label=left T-DNA border
                     /note="left T-DNA border"
     primer_bind     3867..3883
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3885..4105)
                     /regulatory_class="terminator"
                     /note="nopaline synthase (nos) terminator"
     terminator      3896..4148
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4151..4945)
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /note="kanamycin resistance"
                     /protein_id="AUG68174.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(4946..6272)
                     /regulatory_class="promoter"
                     /note="Arabidopsis thaliana UBIQUITIN10 (At4g05320) 
                     promoter plus 5-UTR"
     terminator      6310..6562
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(6313..9672)
                     /note="Overexpression cassette of PtGAUT12 
                     (Potri.001G416800)"
     regulatory      complement(6313..6512)
                     /regulatory_class="terminator"
                     /note="nopaline synthase (nos) terminator"
     protein_bind    6597..6621
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(6598..6618)
                     /label=attB2
                     /note="attB2"
     CDS             complement(6646..8247)
                     /codon_start=1
                     /product="galacturonosyltransferase 12"
                     /label=galacturonosyltransferase 12
                     /note="PtGAUT12.1 (Potri.001G416800); GAUT12"
                     /protein_id="AUG68175.1"
                     /translation="MQLHISPSLRHVTVFPGKGVREFIKVRVGARRVSYRMLFYSLLFF
                     TFLLRFVFVLSTVDSIDGETKCSTLGCLGKRLGPRILGRRLDSAVPEVMFQVLEQPLGN
                     DELKGRSDIPQTLEEFMDEVKNTRLDAKTFALKLREMVTLLEQRTRNAKIQEYLYRHVA
                     SSSIPKQLHCLALRLASEHSTNAAARLQLPLPELVPALVDNTYFHFVLASDNVLAAAVV
                     ANSLVQNALRPQKFVLHIITDRKTYSPMQAWFSLHPLAPAIIEVKALHHFDWFAKGKVP
                     VMEAMEKDQRVRSQFRGGSSAIVANNTEKPHIIAAKLQTLSPKYNSVMNHIRIHLPELF
                     PSLNKVVFLDDDIVVQSDLSPLWDIDMNGKVNGAVETCRGEDKFVMSKKLKSYLNFSHP
                     LISENFKPNECAWAYGMNIFDLEAWRKTNISTTYHHWVEENLKSDLSLWQLGTLPPGLI
                     AFHGHVHVIDPFWHMLGLGYQENTSLADAETAGVIHFNGRAKPWLDIAFPQLRPLWAKY
                     INFSDKFIKGCHIRPS"
     protein_bind    complement(8274..8298)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(8277..8297)
                     /label=attB1
                     /note="attB1"
     regulatory      complement(8320..9672)
                     /regulatory_class="promoter"
                     /note="Arabidopsis thaliana UBIQUITIN3 (At5g03240) promoter
                     (- intron)"
     primer_bind     complement(9676..9692)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9745..9769
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    complement(9746..9769)
                     /label=right T-DNA border
                     /note="right T-DNA border"