LOCUS       Exported                5405 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Moss transformation vector p3xHA NPTII DNA, complete sequence.
ACCESSION   LC101762
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5405)
  AUTHORS   Nishiyama T, Cheng C, Tamada Y, Kurata T, Hasebe M.
  TITLE     Moss transformation vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5405)
  AUTHORS   Nishiyama T, Cheng C, Tamada Y, Kurata T, Hasebe M.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-NOV-2015) Contact:Yosuke Tamada National Institute for
            Basic Biology, Division of Evolutionary Biology; 38 Nishigonaka, 
            Myodaiji, Okazaki, Aichi 444-8585, Japan URL 
            :http://www.nibb.ac.jp/evodevo/
REFERENCE   3  (bases 1 to 5405)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5405
                     /organism="Moss transformation vector p3xHA NPTII"
                     /mol_type="other DNA"
                     /db_xref="taxon:1765186"
     misc_feature    1..652
                     /label=pBluescript II KS +
                     /note="pBluescript II KS +"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             701..802
                     /codon_start=1
                     /gene="3xHA"
                     /product="Three tandem repeats of hemagglutinin tag"
                     /label=3xHA
                     /note="3xHA without start methionine"
                     /protein_id="BAU09067.1"
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYAVYK"
     gene            701..802
                     /gene="3xHA"
                     /label=3xHA
     CDS             701..790
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_recomb     822..855
                     /label=loxP
                     /note="loxP"
     protein_bind    822..855
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    861..1111
                     /label=nos terminator
                     /note="nos terminator"
     regulatory      1147..1538
                     /regulatory_class="promoter"
                     /note="Modified cauliflower mosaic virus 35S promoter"
     promoter        1493..1539
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1562..2356
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /protein_id="BAU09068.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            1562..2356
                     /gene="nptII"
                     /label=nptII
     misc_feature    2385..3084
                     /label=Cauliflower mosaic virus 35S polyA signal
                     /note="Cauliflower mosaic virus 35S polyA signal"
     polyA_signal    2426..2602
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3091..3124
                     /function="Recombination site"
                     /label=loxP
                     /note="loxP"
     protein_bind    3091..3124
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(3217..3235)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3256..3272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3280..3296
                     /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(3304..3334)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3349..3370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    3544..5405
                     /label=pBluescript II KS +
                     /note="pBluescript II KS +"
     rep_origin      complement(3658..4246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4417..5277)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5278..5382)
                     /gene="bla"
                     /label=AmpR promoter