LOCUS       Exported               13372 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCTAGII-GUSPlus, complete sequence.
ACCESSION   MG818373
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13372)
  AUTHORS   Thomson JG.
  TITLE     pCTAGII binary vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13372)
  AUTHORS   Thomson JG.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-JAN-2018) Crop Improvement and Genetics Reseach Unit, 
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   3  (bases 1 to 13372)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..13372
                     /organism="Cloning vector pCTAGII-GUSPlus"
                     /mol_type="other DNA"
                     /db_xref="taxon:2447765"
     misc_feature    1..25
                     /label=Agrobacterium T-DNA right border
                     /note="Agrobacterium T-DNA right border"
     misc_feature    35..352
                     /label=Zea mays Ds transposon
                     /note="Zea mays Ds transposon"
     misc_feature    360..393
                     /label=loxP recognition site for Cre recombinase
                     /note="loxP recognition site for Cre recombinase"
     protein_bind    360..393
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    435..540
                     /label=MRS recognition site for ParA recombinase
                     /note="MRS recognition site for ParA recombinase"
     misc_feature    574..692
                     /label=RS2 recognition site for cinH recombinase
                     /note="RS2 recognition site for cinH recombinase"
     regulatory      complement(760..1012)
                     /regulatory_class="terminator"
                     /note="nos terminator"
     terminator      760..1012
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1073..3169)
                     /codon_start=1
                     /gene="codA-nptII"
                     /product="bifunctional CodA-NPTII fusion protein"
                     /function="confers sensitivity to 5-fluorocytosine (5-FC) 
                     and resistance to kanamycin in plants"
                     /label=codA-nptII
                     /note="cytosine deaminase/neomycin phosphotransferase II 
                     fusion protein; translational fusion between E. coli 
                     cytosine deaminase (codA) and neomycin phosphotransferase 
                     II (nptII) genes"
                     /protein_id="AYG86907.1"
                     /translation="MSNNALQTIINARLPGEEGLWQIHLQDGKISAIDAQSGVMPITEN
                     SLDAEQGLVIPPFVEPHIHLDTTQTAGQPNWNQSGTLFEGIERWAERKALLTHDDVKQR
                     AWQTLKWQIANGIQHVRTHVDVSDATLTALKAMLEVKQEVAPWIDLQIVAFPQEGILSY
                     PNGEALLEEALRLGADVVGAIPHFEFTREYGVESLHKTFALAQKYDRLIDVHCDEIDDE
                     QSRFVETVAALAHHEGMGARVTASHTTAMHSYNGAYTSRLFRLLKMSGINFVANPLVNI
                     HLQGRFDTYPKRRGITRVKEMLESGINVCFGHDDVFDPWYPLGTANMLQVLHMGLHVCQ
                     LMGYGQINDGLNLITHHSARTLNLQDYGIAAGNSANLIILPAENGFDALRRQVPVRYSV
                     RGGKVIASTQPAQTTVYLEQPEAIDYKRAAAGGGMGIEQDGLHAGSPAAWVERLFGYDW
                     AQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDV
                     VTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIE
                     RARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENG
                     RFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLL
                     DEFF"
     gene            complement(1073..3169)
                     /gene="codA-nptII"
                     /label=codA-nptII
     CDS             complement(1073..1870)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1889..3169)
                     /codon_start=1
                     /gene="codA"
                     /product="E. coli cytosine deaminase"
                     /label=codA
                     /note="confers sensitivity to 5-fluorocytosine"
                     /translation="MSNNALQTIINARLPGEEGLWQIHLQDGKISAIDAQSGVMPITEN
                     SLDAEQGLVIPPFVEPHIHLDTTQTAGQPNWNQSGTLFEGIERWAERKALLTHDDVKQR
                     AWQTLKWQIANGIQHVRTHVDVSDATLTALKAMLEVKQEVAPWIDLQIVAFPQEGILSY
                     PNGEALLEEALRLGADVVGAIPHFEFTREYGVESLHKTFALAQKYDRLIDVHCDEIDDE
                     QSRFVETVAALAHHEGMGARVTASHTTAMHSYNGAYTSRLFRLLKMSGINFVANPLVNI
                     HLQGRFDTYPKRRGITRVKEMLESGINVCFGHDDVFDPWYPLGTANMLQVLHMGLHVCQ
                     LMGYGQINDGLNLITHHSARTLNLQDYGIAAGNSANLIILPAENGFDALRRQVPVRYSV
                     RGGKVIASTQPAQTTVYLEQPEAIDYKR"
     regulatory      complement(3177..3978)
                     /regulatory_class="promoter"
                     /note="Arabidopsis ubiquitin 10 promoter and 5 intron"
     intron          complement(3193..3480)
                     /note="AtUbi10 5 intron"
     CDS             join(4094..4108,4299..6146)
                     /codon_start=1
                     /gene="GUSPlus"
                     /product="beta-glucuronidase"
                     /label=GUSPlus
                     /note="reporter gene in plants that encodes a protein that 
                     enzymatically converts a colorless X-gluc substrate into a 
                     blue product"
                     /protein_id="AYG86908.1"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKNASHHHHHHV"
     gene            4094..6146
                     /gene="GUSPlus"
                     /label=GUSPlus
     CDS             join(4094..4108,4299..6116)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=GUSPlus(TM)
                     /note="codon-optimized Staphylococcus gusA gene with a 
                     catalase intron to ensure expression in plants but not 
                     bacteria"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     intron          4109..4298
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             6123..6140
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      6172..6424
                     /regulatory_class="terminator"
                     /note="nos terminator"
     terminator      6172..6424
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6570..6608
                     /label=PhiC31 attP recognition site
                     /note="PhiC31 attP recognition site"
     misc_feature    6628..6881
                     /label=Zea mays Ds transposon
                     /note="Zea mays Ds transposon"
     misc_feature    6969..6993
                     /label=Agrobacterium T-DNA left border
                     /note="Agrobacterium T-DNA left border"
     CDS             7418..8212
                     /codon_start=1
                     /gene="APH(3)-IIIa"
                     /product="aminoglycoside phosphotransferase APH(3)-IIIa"
                     /function="confers kanamycin resistance"
                     /label=APH(3)-IIIa
                     /protein_id="AYG86909.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            7418..8212
                     /gene="APH(3)-IIIa"
                     /label=APH(3)-IIIa
     rep_origin      8299..8887
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     misc_feature    9073..9213
                     /note="bom; basis of mobility region from pBR322"
     rep_origin      9557..9751
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9817..10884)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein A; from Pseudomonas plasmid 
                     pVS1"
                     /protein_id="AYG86910.1"
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     gene            complement(9817..10884)
                     /gene="repA"
                     /label=repA
     CDS             complement(11318..11947)
                     /codon_start=1
                     /gene="staA"
                     /product="StaA"
                     /label=staA
                     /note="stability protein; from Pseudomonas plasmid pVS1"
                     /protein_id="AYG86911.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     gene            complement(11318..11947)
                     /gene="staA"
                     /label=staA
     misc_feature    13247..13271
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(13327..13344)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"