LOCUS       Exported               12362 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Standard reference vector pMCS, complete sequence.
ACCESSION   KJ701603
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12362)
  AUTHORS   Xi J, Wang X, Dong L, Dong Y, Su Y, Tang Q, Wang Z.
  TITLE     Construction of a standard reference plasmid containing seven target
            genes for the detection of transgenic cotton
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12362)
  AUTHORS   Xi J, Wang X, Dong L, Dong Y, Su Y, Tang Q, Wang Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-APR-2014) Biosafet Lab, Biotechnology Research 
            Institute, No. 12 Zhongguancun South Street, Beijing 100081, China
REFERENCE   3  (bases 1 to 12362)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12362
                     /organism="standard reference vector pMCS"
                     /mol_type="other DNA"
                     /note="constructed based on pCamBIA2300 described in INSDC 
                     Accession Number AF234315"
                     /db_xref="taxon:1488197"
     misc_feature    complement(980..1980)
                     /label=STA region from pVS1 plasmid
                     /note="STA region from pVS1 plasmid"
     CDS             1219..1848
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2282..3349
                     /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      complement(2573..3573)
                     /direction=LEFT
                     /note="pVS1-REP; replication origin from pVS1"
     rep_origin      3415..3609
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    3953..4093
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4279..4867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(4383..4663)
                     /direction=LEFT
                     /label=pBR322 origin of replication
                     /note="pBR322 origin of replication"
     CDS             complement(4954..5748)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside phosphotransferase"
                     /function="kanamycin resistance"
                     /label=aadA
                     /protein_id="AII71481.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(4954..5748)
                     /gene="aadA"
                     /label=aadA
                     /note="amplified from pIG121Hm"
     misc_feature    6173..6197
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    6275..6449
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(6506..7303)
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /function="kanamycin resistance"
                     /label=nptII
                     /protein_id="AII71482.1"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(6506..7303)
                     /gene="nptII"
                     /label=nptII
     regulatory      complement(7334..8114)
                     /regulatory_class="promoter"
                     /note="from CaMV35S2, duplicated"
     promoter        complement(7366..8043)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    8234..8255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8270..8300
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8308..8324
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8332..8348
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8364..8633
                     /codon_start=1
                     /gene="cptI"
                     /product="cowpea trypsin inhibitor"
                     /label=cptI
                     /protein_id="AII71483.1"
                     /translation="MRMDLKHLGSNHHDDSSDEPSESSEPCCDSCICTKSIPPQCHCTD
                     IRWNSCHSACKSCMCTRSMPGKCRCLDIADFCYKPCKSRDEDDE"
     gene            8364..8633
                     /gene="cptI"
                     /label=cptI
     CDS             complement(8551..9579)
                     /codon_start=1
                     /gene="Cry1Ab/1Ac"
                     /product="Cry1Ab/Cry1Ac fusion protein"
                     /label=Cry1Ab/1Ac
                     /note="Bt protein"
                     /protein_id="AII71484.1"
                     /translation="NCLSNPEVVVLGGERIETGYTPIDISLSLTQFLLSEFVPGAGFVL
                     GPVDIIWGIFGPSQWDAFLVQIEQLINQRIEEFARNQAISRLEGLSNLYQIYAESFREW
                     EADPTNPALREEMRIQFNDMNSALTTAIPLFAVQNYQVPLLSVYVQAANLHLSVLRDVS
                     VFGQRWGFDAATINSRYNDLTRLIGNYTDHAVRWYNTGLERVWGPDSRDWIRYNQFRRE
                     LTLTVLDIVSLFPNYDSRTYPIRTVSQLTREIYTNPVLENFDGSFRGSAQGIEGSIRSP
                     HLMDILNSITIYTDAHRGEYYWSGHQIMASPVGFIIRRLRHEICTVCSRSPRCPGNGIY
                     PA"
     gene            complement(8551..9579)
                     /gene="Cry1Ab/1Ac"
                     /label=Cry1Ab/1Ac
     CDS             9595..10962
                     /codon_start=1
                     /gene="aroA"
                     /product="5-enolpyruvulshikimate-3-phosphate synthase"
                     /label=aroA
                     /note="cp4-epsps"
                     /protein_id="AII71485.1"
                     /translation="MAHGASSRPATARKSSGLSGTVRIPGDKSISHRSFMFGGLASGET
                     RITGLLEGEDVINTGKAMQAMGARIRKEGDTWIIDGVGNGGLLAPEAPLDFGNAATGCR
                     LTMGLVGVYDFDSTFIGDASLTKRPMGRVLNPLREMGVQVKSEDGDRLPVTLRGPKTPT
                     PITYRVPMASAQVKSAVLLAGLNTPGITTVIEPIMTRDHTEKMLQGFGANLTVETDADG
                     VRTIRLEGRGKLTGQVIDVPGDPSSTAFPLVAALLVPGSDVTILNVLMNPTRTGLILTL
                     QEMGADIEVINPRLAGGEDVADLRVRSSTLKGVTVPEDRAPSMIDEYPILAVAAAFAEG
                     ATVMNGLEELRVKESDRLSAVANGLKLNGVDCDEGETSLVVRGRPDGKGLGNASGAAVA
                     THLDHRIAMSFLVMGLVSENPVTVDDATMIATSFPEFMDLMAGLGAKIELSDTKAA"
     gene            9595..10962
                     /gene="aroA"
                     /label=aroA
     regulatory      10963..11180
                     /regulatory_class="terminator"
                     /note="NOS"
     CDS             join(11182..11296,11671..12064)
                     /codon_start=1
                     /gene="SadI"
                     /product="AE9 stearoyl-ACP desaturase"
                     /label=SadI
                     /note="from cotton K312"
                     /protein_id="AII71486.1"
                     /translation="STMPSPRNLRSSLALLFHQRPPLDLPSFP*SPPFLLAPKRLGI*K
                     SLSRLQRRCLFRSPTPCRLTRLRSLNLWRAGLRTTF*LTSNQLRNVGNPPTFFQILILM
                     DFMSKSKSLGKGQRRSQMITL*FWLVI*SPRKPFQLIKQCLIPWMELVMRQVLASLALA
                     VVLQRR"
     gene            11182..12064
                     /gene="SadI"
                     /label=SadI
     primer_bind     complement(12038..12054)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    12257..12281
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"