LOCUS       Exported               14105 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMCG161, complete sequence.
ACCESSION   AY572837
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14105)
  AUTHORS   Sidorenko L, McGinnis K, Jorgensen R, Chandler V.
  TITLE     Vector for silencing of maize chromatin genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14105)
  AUTHORS   Sidorenko L, McGinnis K, Jorgensen R, Chandler V.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-MAR-2004) Plant Sciences, University of Arizona, 303 
            Forbes Hall, Tucson, AZ 85721, USA
REFERENCE   3  (bases 1 to 14105)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14105
                     /organism="Cloning vector pMCG161"
                     /mol_type="other DNA"
                     /label=used for production of RNAi lines
                     /note="used for production of RNAi lines"
                     /db_xref="taxon:269149"
     regulatory      34..269
                     /regulatory_class="polyA_signal_sequence"
                     /note="nos 3"
     CDS             complement(294..845)
                     /codon_start=1
                     /gene="bar"
                     /product="phosphinothricin acetyl transferase"
                     /label=bar
                     /note="resistance to herbicide BASTA"
                     /protein_id="AAS79014.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            complement(294..845)
                     /gene="bar"
                     /label=bar
     promoter        complement(875..2864)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     misc_feature    879..2864
                     /label=maize ubiquitin intron 1 and promoter
                     /note="maize ubiquitin intron 1 and promoter"
     misc_feature    2882..3439
                     /label=small subunit ribosomal RNA (ssu) 3
                     /note="small subunit ribosomal RNA (ssu) 3"
     regulatory      3476..4807
                     /regulatory_class="promoter"
                     /note="Cauliflower mosaic virus 35S promoter"
     promoter        4457..4804
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     intron          4837..5402
                     /number=1
                     /note="maize adh1"
     misc_feature    5422..5441
                     /note="multiple cloning site; includes AscI, EcoRV, RsrII, 
                     and AvrII; used for cloning first inverted repeat, forward 
                     orientation"
     intron          5443..6580
                     /number=1
                     /note="rice Nipponbare waxy-a"
     misc_feature    6584..6604
                     /note="multiple cloning site; includes SgfI, SmaI, and 
                     SpeI; used for cloning second inverted repeat, reverse 
                     orientation"
     regulatory      6620..7332
                     /regulatory_class="polyA_signal_sequence"
                     /note="ocs 3"
     terminator      6623..7330
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     complement(7355..7371)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7574..7598
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             8898..9527
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9961..11028
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      11094..11288
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    11632..11772
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(11958..12546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(12817..13476)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acyl transferase"
                     /label=cat
                     /note="chloramphenicol resistance"
                     /protein_id="AAS79015.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(12817..13476)
                     /gene="cat"
                     /label=cat
     misc_feature    14004..14028
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"