LOCUS       Exported                6567 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Moss transformation vector pTFH22.4 DNA, complete sequence.
ACCESSION   AB758445
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6567)
  AUTHORS   Sakakibara K, Reisewitz P, Aoyama T, Friedrich T, Ando S, Sato Y, 
            Nishiyama T, Tamada Y, Hiwatashi Y, Kurata T, Ishikawa M, Deguchi H,
            Rensing SA, Werr W, Murata T, Hasebe M, Laux T.
  TITLE     WOX13-like genes are required for the initiation of apical stem 
            cells from differentiated leaf cells in the moss Physcomitrella 
            patens
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6567)
  AUTHORS   Sakakibara K, Fujita T, Hasebe M.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-OCT-2012) Contact:Keiko Sakakibara Graduate School of 
            Science, Hiroshima University, Department of Biological Science; 
            1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8526, Japan
REFERENCE   3  (bases 1 to 6567)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6567
                     /organism="Moss transformation vector pTFH22.4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1263457"
     rep_origin      28..616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    904..925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        940..970
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    978..994
                     /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     1002..1018
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1045..2458
                     /label=Pact=rice actin promoter
                     /note="Pact=rice actin promoter"
     CDS             1898..1915
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2450..2976
                     /label=TrbcS
                     /note="TrbcS"
     misc_feature    2461..2496
                     /note="SfiI Multi cloning site (SfiI, AscI, SmaI, ApaI, 
                     SfiI)"
     misc_feature    2989..4613
                     /label=nptII expression cassette
                     /note="nptII expression cassette"
     promoter        3354..3400
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             3423..4217
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      4656..5530
                     /regulatory_class="promoter"
                     /note="Cauliflower Mosaic Virus 35S promoter"
     promoter        5143..5488
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             5536..6255
                     /codon_start=1
                     /product="modified green fluorescent protein"
                     /label=modified green fluorescent protein
                     /protein_id="BAN16591.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     terminator      6263..6515
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"