LOCUS       Exported               11439 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCTAG6, complete sequence.
ACCESSION   MG836292
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11439)
  AUTHORS   Thomson JG.
  TITLE     pCTAG6
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11439)
  AUTHORS   Thomson JG.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JAN-2018) Crop Improvement and Genetics Reseach Unit, 
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   3  (bases 1 to 11439)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11439
                     /organism="Cloning vector pCTAG6"
                     /mol_type="other DNA"
                     /db_xref="taxon:2447764"
     misc_feature    1..25
                     /label=Agrobacterium T-DNA right border
                     /note="Agrobacterium T-DNA right border"
     misc_feature    35..351
                     /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(726..1108)
                     /regulatory_class="terminator"
                     /note="Arabidopsis ubiquitin 10 transcription terminator"
     CDS             complement(1121..3217)
                     /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="AYG86912.1"
                     /translation="MSNNALQTIINARLPGEEGLWQIHLQDGKISAIDAQSGVMPITEN
                     SLDAEQGLVIPPFVEPHIHLDTTQTAGQPNWNQSGTLFEGIERWAERKALLTHDDVKQR
                     AWQTLKWQIANGIQHVRTHVDVSDATLTALKAMLEVKQEVAPWIDLQIVAFPQEGILSY
                     PNGEALLEEALRLGADVVGAIPHFEFTREYGVESLHKTFALAQKYDRLIDVHCDEIDDE
                     QSRFVETVAALAHHEGMGARVTASHTTAMHSYNGAYTSRLFRLLKMSGINFVANPLVNI
                     HLQGRFDTYPKRRGITRVKEMLESGINVCFGHDDVFDPWYPLGTANMLQVLHMGLHVCQ
                     LMGYGQINDGLNLITHHSARTLNLQDYGIAAGNSANLIILPAENGFDALRRQVPVRYSV
                     RGGKVIASTQPAQTTVYLEQPEAIDYKRAAAGGGMGIEQDGLHAGSPAAWVERLFGYDW
                     AQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDV
                     VTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIE
                     RARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENG
                     RFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLL
                     DEFF"
     gene            complement(1121..3217)
                     /gene="codA-nptII"
                     /label=codA-nptII
     CDS             complement(1121..1918)
                     /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(1937..3217)
                     /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(3229..4039)
                     /regulatory_class="promoter"
                     /note="Arabidopsis Ubiquitin 10 promoter and 5 intron"
     intron          complement(3245..3532)
                     /note="Arabidopsis 5 enhancing intron"
     misc_recomb     4055..4106
                     /note="attB recognition site for Bxb1 site-specific 
                     recombinase"
     misc_recomb     4166..4222
                     /note="attP recognition site for phiC31 site-specific 
                     recombinase"
     misc_feature    4232..4501
                     /label=Zea mays Ds transposon
                     /note="Zea mays Ds transposon"
     misc_feature    5035..5059
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5484..6278
                     /codon_start=1
                     /gene="APH(3)-IIIa"
                     /product="aminoglycoside phosphotransferase APH(3)-IIIa"
                     /function="confers kanamycin resistance"
                     /label=APH(3)-IIIa
                     /protein_id="AYG86913.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            5484..6278
                     /gene="APH(3)-IIIa"
                     /label=APH(3)-IIIa
     rep_origin      6365..6953
                     /note="/function=ColE1 origin of replication"
     misc_feature    7139..7279
                     /note="bom; basis of mobility region from pBR322"
     rep_origin      7623..7817
                     /note="/function=pVS1 oriV broad host range origin of 
                     replication"
     CDS             complement(7883..8950)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein A; from Pseudomonas plasmid 
                     pVS1"
                     /protein_id="AYG86914.1"
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     gene            complement(7883..8950)
                     /gene="repA"
                     /label=repA
     CDS             complement(9384..10013)
                     /codon_start=1
                     /gene="staA"
                     /product="StaA"
                     /label=staA
                     /note="stability protein; from Pseudomonas plasmid pVS1"
                     /protein_id="AYG86915.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     gene            complement(9384..10013)
                     /gene="staA"
                     /label=staA
     misc_feature    11313..11337
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(11393..11410)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"