LOCUS       Exported                6587 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBS414V, complete sequence.
ACCESSION   AY289542
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6587)
  AUTHORS   Tooley AJ, Cai YA, Glazer AN.
  TITLE     Biosynthesis of a fluorescent cyanobacterial C-phycocyanin 
            holo-alpha subunit in a heterologous host
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 98 (19), 10560-10565 (2001)
  PUBMED    11553806
REFERENCE   2  (bases 1 to 6587)
  AUTHORS   Tooley AJ, Cai YA, Glazer AN.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2003) Molecular and Cell Biology, University of 
            California, Berkeley, 142 LSA no. 3200, Berkeley, CA 94720-3200, USA
REFERENCE   3  (bases 1 to 6587)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6587
                     /organism="Expression vector pBS414V"
                     /mol_type="other DNA"
                     /db_xref="taxon:231029"
     gene            10..195
                     /gene="rop"
                     /label=rop
                     /note="coding region not determined; repressor of primer 
                     protein variant; regulation of plasmid replication; 
                     maintains medium plasmid copy number"
     misc_feature    37..1286
                     /note="contains oriV and oriT regions; sequence derived 
                     from plasmid pBR322 of Genbank Accession Number J01749"
     misc_feature    297..439
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(625..1213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1497..2285
                     /codon_start=1
                     /gene="aadA"
                     /product="AadA"
                     /label=aadA
                     /note="confers resistance to streptomycin and to 
                     spectinomycin via inactivation of the drugs by specific 
                     adenylylation"
                     /protein_id="AAP43512.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            1497..2285
                     /gene="aadA"
                     /label=aadA
     CDS             2506..3588
                     /codon_start=1
                     /gene="lacIq"
                     /product="LacI"
                     /label=lacIq
                     /note="lac repressor LacI protein; represses transcription 
                     by binding to the lac promoter/operator region"
                     /protein_id="AAP43513.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            2506..3588
                     /gene="lacIq"
                     /label=lacIq
     regulatory      3820..3825
                     /regulatory_class="minus_35_signal"
                     /note="trc promoter"
     regulatory      3843..3849
                     /regulatory_class="minus_10_signal"
                     /note="trc promoter"
     protein_bind    3853..3878
                     /label=LacI binding site
                     /bound_moiety="LacI"
     misc_feature    3855..3856
                     /note="transcription initiation site from the trc promoter"
     protein_bind    3857..3873
                     /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)."
     gene            3882..4454
                     /gene="cpcA"
                     /label=cpcA
     regulatory      3882..3887
                     /regulatory_class="ribosome_binding_site"
                     /gene="cpcA"
     CDS             3894..4454
                     /codon_start=1
                     /gene="cpcA"
                     /product="C-phycocyanin alpha subunit apoprotein"
                     /label=cpcA
                     /note="from Synechocystis sp. PCC 6803; fused at the 
                     N-terminus to a 24 amino acid sequence with six-His tag"
                     /protein_id="AAP43514.1"
                     /translation="MGHHHHHHDYDIPTTENLYFQGAHMKTPLTEAVSTADSQGRFLSS
                     TELQIAFGRLRQANAGLQAAKALTDNAQSLVNGAAQAVYNKFPYTTQTQGNNFAADQRG
                     KDKCARDIGYYLRIVTYCLVAGGTGPLDEYLIAGIDEINRTFDLSPSWYVEALKYIKAN
                     HGLSGDARDEANSYLDYAINALS"
     CDS             3900..3917
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3939..3959
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     gene            4483..5313
                     /gene="cpcE"
                     /label=cpcE
     regulatory      4483..4488
                     /regulatory_class="ribosome_binding_site"
                     /gene="cpcE"
     CDS             4495..5313
                     /codon_start=1
                     /gene="cpcE"
                     /product="CpcE"
                     /label=cpcE
                     /note="required for the covalent attachment of 
                     phycocyanobilin to the alpha subunit apoprotein of 
                     C-phycocyanin"
                     /protein_id="AAP43515.1"
                     /translation="MSEPNLNPAYTLDQAIANLQQTEDASARYYAAWWIGRFRAAQPET
                     IAALLVALEDETDRSPDGGYPLRRNAAKALGKLGDRQVVPALIKALECEDYYVRESAAQ
                     ALEGLGDARAMAPLMAKLTGGLAAAQLVEGKPHLAQPYEAIIEALGTLQAVESIGLIEP
                     FLEHFSPKVQYAAARALFQLTGDNRYGDLLITALGGTDLQLRRSAMMDLGATGYLPGAQ
                     AIAKAFAENSLKLIALRDLWATHRQRQASSESKALSPASRQILELMDSLL"
     gene            5330..5988
                     /gene="cpcF"
                     /label=cpcF
     regulatory      5330..5335
                     /regulatory_class="ribosome_binding_site"
                     /gene="cpcF"
     CDS             5344..5988
                     /codon_start=1
                     /gene="cpcF"
                     /product="CpcF"
                     /label=cpcF
                     /note="required for the covalent attachment of 
                     phycocyanobilin to the alpha subunit apoprotein of 
                     C-phycocyanin"
                     /protein_id="AAP43516.1"
                     /translation="MEGNSVVTPEIERLIQAVETADSAAKLVGAVRALAATRSPLAVPQ
                     LTTVLRYNNPGAAVAAVDGLIQIGDAAMTHLLANMDGYNYGARAWATRACAGIGDPRAL
                     ALLQEAALTDFALSVRRAAAKGLGFLRWQSLPQEEQETVQKAIYDTLIQVCEDPEWVVR
                     YGAIAGLENLAKQAQHYRQPLKDFLQSFVEQEPEAIVGERILWTLENIGPI"
     rRNA            6134..6548
                     /gene="rrnB"
                     /product="5S ribosomal RNA"
                     /label=rrnB
                     /note="contains two strong, bidirectional, rho-independent 
                     transcription terminator sequences"
     gene            6134..6548
                     /gene="rrnB"
                     /label=rrnB
     terminator      6324..6410
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6502..6529
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"