LOCUS       Exported                4153 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Plant expression vector pPILY, complete sequence.
ACCESSION   AY720433
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4153)
  AUTHORS   Ferrando A, Farras R, Jasik J, Schell J, Koncz C.
  TITLE     Intron-tagged epitope: a tool for facile detection and purification 
            of proteins expressed in Agrobacterium-transformed plant cells
  JOURNAL   Plant J. 22 (6), 553-560 (2000)
  PUBMED    10886775
REFERENCE   2  (bases 1 to 4153)
  AUTHORS   Berendzen KW, Koncz C.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-AUG-2004) Development, Max-Planck Institute for Plant 
            Breeding Research, Carl-von-Linne-Weg 10, Cologne D-50829, Germany
REFERENCE   3  (bases 1 to 4153)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4153
                     /organism="Plant expression vector pPILY"
                     /mol_type="other DNA"
                     /db_xref="taxon:292598"
     misc_feature    1..18
                     /note="Tth111I, NotI, XhoI, KpnI; MCS promoter"
     regulatory      19..758
                     /regulatory_class="promoter"
                     /note="duplicated 35S CaMV (Cabb B-JI isolate); 35SS; 
                     overexpression in plants"
     promoter        409..754
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    759..793
                     /note="HindIII, NcoI, SmaI, XbaI, BglII; MCS upstream"
     misc_feature    794..805
                     /label=N-term HA epitope
                     /note="N-term HA epitope"
     intron          806..994
                     /note="PIV2 optimized; prevents detection of epitope tag in
                     bacteria"
     misc_feature    995..1009
                     /label=C-term HA epitope
                     /note="C-term HA epitope"
     misc_feature    1010..1037
                     /note="SalI, BamHI, PstI, EcoRI; MCS downstream"
     regulatory      1038..1290
                     /regulatory_class="terminator"
                     /note="polyadenylation signal from nopaline synthase gene 
                     of the Agrobacterium Ti plasmid; NOS terminator"
     misc_feature    1291..1360
                     /note="KpnI, SacI, NotI, SacI, T7, SfiI; MCS terminator"
     promoter        complement(1322..1340)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(1361..1589)
                     /label=LacI
                     /note="LacI"
     primer_bind     complement(1370..1386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1394..1410
                     /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(1418..1448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1463..1484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1772..2360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2531..3391)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="resistance to ampcillin"
                     /protein_id="AAU21232.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2531..3391)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(3392..3496)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3523..3978
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4119..4135
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"