LOCUS       Exported                4976 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pAT101, complete sequence.
ACCESSION   AY302760
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4976)
  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 4976)
  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   3  (bases 1 to 4976)
  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   4  (bases 1 to 4976)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4976
                     /organism="Expression vector pAT101"
                     /plasmid="pAT101"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:231720"
     misc_feature    1..2773
                     /note="vector backbone sequence derived from plasmid 
                     pACYC177 of Genbank accession number X06402"
     rep_origin      54..883
                     /label=p15A origin of replication
                     /note="p15A origin of replication"
     rep_origin      71..616
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             1211..2026
                     /codon_start=1
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aminoglycoside 3-phosphotransferase
                     /note="confers resistance to kanamycin"
                     /protein_id="AAP45706.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      2790..2795
                     /regulatory_class="minus_35_signal"
                     /note="trc promoter"
     regulatory      2813..2819
                     /regulatory_class="minus_10_signal"
                     /note="trc promoter"
     protein_bind    2823..2848
                     /label=LacI binding site
                     /bound_moiety="LacI"
     misc_feature    2825..2826
                     /label=transcription initiation site from trc promoter
                     /note="transcription initiation site from trc promoter"
     protein_bind    2827..2843
                     /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            2852..3658
                     /gene="hox1"
                     /label=hox1
                     /note="sll1184"
     regulatory      2852..2857
                     /regulatory_class="ribosome_binding_site"
                     /gene="hox1"
     CDS             2864..3658
                     /codon_start=1
                     /gene="hox1"
                     /product="heme oxygenase 1"
                     /label=hox1
                     /note="from Synechocystis sp. PCC 6803; consists of HO1 
                     fused at the N terminus to a 24-amino-acid sequence that 
                     includes a six-His tag; catalyzes the conversion of heme to
                     biliverdin"
                     /protein_id="AAP45707.1"
                     /translation="MGHHHHHHDYDIPTTENLYFQGAHMSVNLASQLREGTKKSHSMAE
                     NVGFVKCFLKGVVEKNSYRKLVGNLYFVYSAMEEEMAKFKDHPILSHIYFPELNRKQSL
                     EQDLQFYYGSNWRQEVKISAAGQAYVDRVRQVAATAPELLVAHSYTRYLGDLSGGQILK
                     KIAQNAMNLHDGGTAFYEFADIDDEKAFKNTYRQAMNDLPIDQATAERIVDEANDAFAM
                     NMKMFNELEGNLIKAIGIMVFNSLTRRRSQGSTEVGLATSEG"
     CDS             2870..2887
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2909..2929
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     gene            3677..4436
                     /gene="pcyA"
                     /label=pcyA
                     /note="slr0116"
     regulatory      3677..3682
                     /regulatory_class="ribosome_binding_site"
                     /gene="pcyA"
     CDS             3690..4436
                     /codon_start=1
                     /gene="pcyA"
                     /product="3Z-phycocyanobilin:ferredoxin oxidoreductase"
                     /label=pcyA
                     /note="PcyA; catalyzes the conversion of biliverdin to 
                     3Z-phycocyanobilin"
                     /protein_id="AAP45708.1"
                     /translation="MAVTDLSLTNSSLMPTLNPMIQQLALAIAASWQSLPLKPYQLPED
                     LGYVEGRLEGEKLVIENRCYQTPQFRKMHLELAKVGKGLDILHCVMFPEPLYGLPLFGC
                     DIVAGPGGVSAAIADLSPTQSDRQLPAAYQKSLAELGQPEFEQQRELPPWGEIFSEYCL
                     FIRPSNVTEEERFVQRVVDFLQIHCHQSIVAEPLSEAQTLEHRQGQIHYCQQQQKNDKT
                     RRVLEKAFGEAWAERYMSQVLFDVIQ"
     rRNA            4523..4937
                     /gene="rrnB"
                     /product="5S ribosomal RNA"
                     /label=rrnB
                     /note="contains two strong, bidirectional, rho-independent 
                     transcription terminator sequences"
     gene            4523..4937
                     /gene="rrnB"
                     /label=rrnB
     terminator      4713..4799
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4891..4918
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"