LOCUS       Exported                6399 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBS437V, complete sequence.
ACCESSION   AY302759
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6399)
  AUTHORS   Tooley AJ, Glazer AN.
  TITLE     Biosynthesis of the cyanobacterial light-harvesting polypeptide 
            phycoerythrocyanin holo-alpha subunit in a heterologous host
  JOURNAL   J. Bacteriol. 184 (17), 4666-4671 (2002)
  PUBMED    12169589
REFERENCE   2  (bases 1 to 6399)
  AUTHORS   Tooley AJ, Glazer AN.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2003) Molecular and Cell Biology, University of 
            California, Berkeley, 142 LSA no. 3200, Berkeley, CA 94720-3200, USA
REFERENCE   3  (bases 1 to 6399)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6399
                     /organism="Expression vector pBS437V"
                     /mol_type="other DNA"
                     /db_xref="taxon:231719"
     gene            10..195
                     /gene="rop"
                     /label=rop
                     /note="coding region not determined; ROP; repressor of 
                     primer protein variant; regulation of plasmid replication; 
                     helps maintain 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="AAP45701.1"
                     /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="AAP45702.1"
                     /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
                     /label=transcription initiation site from trc promoter
                     /note="transcription initiation site from 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="pecA"
                     /label=pecA
     regulatory      3882..3887
                     /regulatory_class="ribosome_binding_site"
                     /gene="pecA"
     CDS             3894..4454
                     /codon_start=1
                     /gene="pecA"
                     /product="phycoerythrocyanin alpha subunit apoprotein"
                     /label=pecA
                     /note="PecA; from Anabaena sp. PCC 7120; consists of PecA 
                     fused at the N-terminus to a 24-amino-acid sequence that 
                     includes a six-His tag"
                     /protein_id="AAP45703.1"
                     /translation="MGHHHHHHDYDIPTTENLYFQGAHMKTPLTEAISAADVRGSYLSN
                     TEMQAVFGRFNRARAGLAAAQAFSNNGKKWAEAAANHVYQKFPYTTQMSGPQYASTPEG
                     KSKCVRDIDHYLRTISYCCVVGGTGPLDEYVVSGLSELNSALGLSPSWYVAALEFVRDN
                     HGLNGDVAGEANIYLNYAINALS"
     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            4487..5259
                     /gene="pecE"
                     /label=pecE
     regulatory      4487..4492
                     /regulatory_class="ribosome_binding_site"
                     /gene="pecE"
     CDS             4498..5259
                     /codon_start=1
                     /gene="pecE"
                     /product="PecE"
                     /label=pecE
                     /note="catalyzes both the covalent attachment of 
                     phycocyanobilin to the phycoerythrocyanin alpha subunit and
                     its concurrent isomerization to phycobiliviolin"
                     /protein_id="AAP45704.1"
                     /translation="MTAEPILSPETAIAALSGEDNQIRYYAAWWLGKHNVQAGFTALCV
                     ALFDERYRIPSGGYPLRRQAARALGLLKNPQAVPALIAALECDEDLRLREAVICSLAAI
                     GDKRAVTPLLNLLQSSQAQPYEALIEALATLQVWSARPQIEPFLQHYSERVQCAAARYM
                     YLLTQESEYIERIVKNLNHDNMYLRWAAVFDLGAVAHQQAVQAILTAQVPNSLKLLNLK
                     RILEAMLNNNSVNHEKAALLFGAIDDLLIQL"
     gene            5268..5800
                     /gene="pecF"
                     /label=pecF
     regulatory      5268..5273
                     /regulatory_class="ribosome_binding_site"
                     /gene="pecF"
     CDS             5279..5800
                     /codon_start=1
                     /gene="pecF"
                     /product="PecF"
                     /label=pecF
                     /note="catalyzes both the covalent attachment of 
                     phycocyanobilin to the phycoerythrocyanin alpha subunit and
                     its concurrent isomerization to phycobiliviolin"
                     /protein_id="AAP45705.1"
                     /translation="MNQASLSVDAITNLIEAFHHHHPAVRSAAVDELIKLGSITVNLLI
                     AAYDDSQDQGFQAQIIQVLAQIGDAKALKLLAEVVGTSVANHCQGNVRRIAARGLGKIA
                     STTSNTEIINNAQEKLIWALLTPEDWGLRYAAAVSLQEIATPKAKAALQQAIAQETDPV
                     VRSRMAIALS"
     rRNA            5946..6360
                     /gene="rrnB"
                     /product="5S ribosomal RNA"
                     /label=rrnB
                     /note="contains two strong, bidirectional, rho-independent 
                     transcription terminator sequences"
     gene            5946..6360
                     /gene="rrnB"
                     /label=rrnB
     terminator      6136..6222
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6314..6341
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"