LOCUS       Exported               11224 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pVC4-bar, complete sequence.
ACCESSION   HQ175992
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11224)
  AUTHORS   Conn SJ, Conn V, Tyerman SD, Kaiser BN, Leigh RA, Gilliham M.
  TITLE     Magnesium transporters, MGT2/MRS2-1 and MGT3/MRS2-5, are important 
            for magnesium partitioning within Arabidopsis thaliana mesophyll 
            vacuoles
  JOURNAL   New Phytol. 190 (3), 583-594 (2011)
  PUBMED    21261624
REFERENCE   2  (bases 1 to 11224)
  AUTHORS   Conn V.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-AUG-2010) Agriculture, Food & Wine, University of 
            Adelaide, PMB1, Glen Osmond, SA 5064, Australia
REFERENCE   3  (bases 1 to 11224)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11224
                     /organism="Cloning vector pVC4-bar"
                     /mol_type="other DNA"
                     /label=derived from Arabidopsis thaliana ecotype Col-0
                     /note="derived from Arabidopsis thaliana ecotype Col-0"
                     /db_xref="taxon:979779"
     misc_feature    82..106
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     regulatory      121..889
                     /regulatory_class="promoter"
                     /note="2X CaMV 35S promoter"
     promoter        526..870
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1078..1094
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     1169..1299
                     /label=AttR1
                     /note="AttR1"
     protein_bind    1175..1299
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1324..1354
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1408..2067
                     /codon_start=1
                     /product="chloramphenicol resistance protein"
                     /label=chloramphenicol resistance protein
                     /protein_id="ADW78951.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2409..2714
                     /codon_start=1
                     /product="CcdB toxicity protein"
                     /label=CcdB toxicity protein
                     /protein_id="ADW78952.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_recomb     2754..2879
                     /label=AttR2
                     /note="AttR2"
     protein_bind    complement(2755..2879)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3191..3207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3215..3231
                     /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)."
     promoter        complement(3239..3269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3284..3305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      3436..4205
                     /regulatory_class="promoter"
                     /note="2X CaMV 35S promoter"
     promoter        3496..4173
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             4218..4769
                     /codon_start=1
                     /product="phosphinothricin acetyltransferase"
                     /label=phosphinothricin acetyltransferase
                     /note="resistance to glyphosate; derived from Streptomyces 
                     hygroscopicus"
                     /protein_id="ADW78953.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    4776..4950
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5028..5052
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5477..6271
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /note="for bacterial selection"
                     /protein_id="ADW78954.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6358..6946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6562..6842
                     /label=pBR322 ori
                     /note="pBR322 ori"
     misc_feature    6982..7242
                     /label=pBR322 bom
                     /note="pBR322 bom"
     misc_feature    7132..7272
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      7616..7810
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      7652..8652
                     /label=pVS1 rep
                     /note="pVS1 rep"
     CDS             complement(7876..8943)
                     /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"
     misc_feature    9245..10245
                     /label=pVS1 sta
                     /note="pVS1 sta"
     CDS             complement(9377..10006)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"