LOCUS       Exported                5756 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Plant expression vector pCB302 LIC GFP, complete sequence.
ACCESSION   KX510271
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  AUTHORS   Zurcher E, Liu J, di Donato M, Geisler M, Muller B.
  TITLE     Plant development regulated by cytokinin sinks
  JOURNAL   Science 353 (6303), 1027-1030 (2016)
  PUBMED    27701112
REFERENCE   2  (bases 1 to 5756)
  AUTHORS   Muller B.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUL-2016) Plant and Microbial Biology, University of 
            Zurich, Zollikerstrasse 107, Zurich, ZH 8008, Switzerland
REFERENCE   3  (bases 1 to 5756)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5756
                     /organism="Plant expression vector pCB302 LIC GFP"
                     /mol_type="other DNA"
                     /note="contains LIC sites for ligation independent cloning 
                     of eGFP translational fusions"
                     /db_xref="taxon:2021232"
     regulatory      6..588
                     /regulatory_class="promoter"
                     /gene="pnos"
     gene            6..588
                     /gene="pnos"
                     /label=pnos
     misc_feature    225..249
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        373..553
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        406..589
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             589..1140
                     /codon_start=1
                     /gene="bar"
                     /product="bar"
                     /label=bar
                     /note="Amp resistance"
                     /protein_id="ASM41179.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            589..1140
                     /gene="bar"
                     /label=bar
     terminator      1172..1424
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_recomb     1433..1462
                     /note="LIC; ligation-independent cloning (LIC) site"
     CDS             1466..2185
                     /codon_start=1
                     /gene="eGFP"
                     /product="eGFP"
                     /label=eGFP
                     /protein_id="ASM41180.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     gene            1466..2185
                     /gene="eGFP"
                     /label=eGFP
     primer_bind     complement(2213..2229)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(2254..2423)
                     /label=T-DNA Right Border
                     /note="T-DNA Right Border"
     misc_feature    2294..2318
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2470..3618)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(3917..4711)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      4923..5554
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5553..5709
                     /label=T-DNA Left Border
                     /note="T-DNA Left Border"
     misc_feature    5632..5656
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     5737..5753
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"