LOCUS       Exported                5667 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Transient expression vector pBI221, complete sequence.
ACCESSION   AF502128
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  AUTHORS   Chen PY, Wang CK, Soong SC, To KY.
  TITLE     Complete sequence of the binary vector pBI121 and its application in
            cloning T-DNA insertion from transgenic plants
  JOURNAL   Mol. Breed. 11, 287-293 (2003)
REFERENCE   2  (bases 1 to 5667)
  AUTHORS   Chen PY, Wang CK, To KY.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-APR-2002) Institute of Bioagricultural Sciences, 
            Academia Sinica, Taipei 11529, Taiwan
REFERENCE   3  (bases 1 to 5667)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5667
                     /organism="Transient expression vector pBI221"
                     /mol_type="genomic DNA"
                     /note="HindIII-EcoRI fragment (3032 bp) carrying CaMV 35S 
                     promoter, GUS coding region and nos terminator was digested
                     from the binary vector pBI121 and ligated with cloning 
                     vector pUC19, which was previously digested with HindIII 
                     and EcoRI; binary vector pBI121 sequence is deposited in 
                     GenBank Accession Number AF485783; plasmids pBI121 and 
                     pBI221 were purchased from Clontech Laboratories, Palo 
                     Alto, CA, USA"
                     /db_xref="taxon:193296"
     regulatory      25..859
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     promoter        512..857
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             896..2707
                     /codon_start=1
                     /gene="gusA"
                     /product="beta-glucuronidase"
                     /label=gusA
                     /protein_id="AAM22211.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            896..2707
                     /gene="gusA"
                     /label=gusA
     regulatory      2778..3030
                     /regulatory_class="terminator"
                     /note="nos terminator"
     terminator      2778..3030
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3039..3055)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3529..3633
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3634..4494
                     /codon_start=1
                     /gene="amp"
                     /product="beta-lactamase"
                     /label=amp
                     /protein_id="AAM22212.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3634..4494
                     /gene="amp"
                     /label=amp
     rep_origin      4642..5285
                     /label=pUC19 plasmid replication origin
                     /note="pUC19 plasmid replication origin"
     rep_origin      4665..5253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5541..5562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5577..5607
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5615..5631
                     /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     5639..5655
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"