LOCUS       Exported                8246 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Chloroplast expression vector pCEV4, complete sequence.
ACCESSION   KY930500
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8246)
  AUTHORS   Chen P-Y., To K-Y.
  TITLE     Construction of chloroplast expression vectors for higher plants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8246)
  AUTHORS   Chen P-Y., To K-Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-APR-2017) Agricultural Biotechnology Research Center, 
            Academia Sinica, No. 128, Academia Road, Section 2, Nankang, Taipei 
            11529, Taiwan
REFERENCE   3  (bases 1 to 8246)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8246
                     /organism="Chloroplast expression vector pCEV4"
                     /mol_type="other DNA"
                     /db_xref="taxon:2127040"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    408..2396
                     /note="Nicotiana benthamiana left targeting region; 
                     contains 16S rRNA and trnI"
     tRNA            join(1609..1645,2362..2396)
                     /product="tRNA-Ile"
     rep_origin      1826..1907
                     /note="chloroplast replication origin; oriA"
     regulatory      2403..2650
                     /regulatory_class="promoter"
                     /note="atpI promoter; PatpI"
     misc_feature    2651..2666
                     /label=cloning site
                     /note="cloning site"
     regulatory      2667..2835
                     /regulatory_class="terminator"
                     /note="rps16 terminator; Trps16"
     regulatory      2842..3060
                     /regulatory_class="promoter"
                     /note="rrn promoter; Prrn"
     CDS             3067..3858
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside 3-adenyltransferase"
                     /label=aadA
                     /note="confers spectinomycin resistance"
                     /protein_id="AVQ67769.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            3067..3858
                     /gene="aadA"
                     /label=aadA
     regulatory      3865..4086
                     /regulatory_class="terminator"
                     /note="psbA terminator; TpsbA"
     misc_feature    4093..5994
                     /note="Nicotiana benthamiana right targeting region; 
                     contains trnA and 23S rRNA"
     tRNA            join(4156..4193,4903..4937)
                     /product="tRNA-Ala"
     primer_bind     complement(6025..6041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6049..6065
                     /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(6073..6103)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6118..6139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6427..7015)
                     /direction=LEFT
                     /label=pUC19 replication origin
                     /note="pUC19 replication origin"
     CDS             complement(7186..8046)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="confers ampicillin resistance"
                     /protein_id="AVQ67770.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7186..8046)
                     /gene="bla"
                     /label=bla
     promoter        complement(8047..8151)
                     /gene="bla"
                     /label=AmpR promoter