LOCUS       Exported                4991 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCDF-AaPKS3, complete sequence.
ACCESSION   MH488926
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  AUTHORS   Kim B, Binkley R, Kim HU, Lee SY.
  TITLE     Metabolic engineering of Escherichia coli for the enhanced 
            production of l-tyrosine
  JOURNAL   Biotechnol. Bioeng. (2018) In press
  PUBMED    30019750
REFERENCE   2  (bases 1 to 4991)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUN-2018) Dept. Chemical and Biomolecular Engineering,
            Korea Advanced Institute of Science and Technology, Daehak-ro 291, 
            Daejeon 34141, South Korea
REFERENCE   3  (bases 1 to 4991)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4991
                     /organism="Cloning vector pCDF-AaPKS3"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:2301141"
     protein_bind    3..27
                     /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)."
     CDS             71..1282
                     /codon_start=1
                     /gene="PKS3"
                     /product="aloesone synthase"
                     /label=PKS3
                     /note="derived from Aloe arborescens"
                     /protein_id="AXN70004.1"
                     /translation="MGSLSDSTPLMKDVQGIRKAQKADGTATVMAIGTAHPPHIISQDS
                     YADFYFRVTNSEHKVELKKKFDRICKKTMIGKRYFNFDEEFLKKYPNITSFDKPSLNDR
                     HDICIPGVPALGAEAAVKAIEEWGRPKSEITHLVFCTSGGVDMPSADFQCAKLLGLRTN
                     VNKYCIYMQGCYAGGTVMRYAKDLAENNRGARVLMVCAELTIIALRGPNDSHIDNAIGN
                     SLFGDGAAALIVGSDPIIGVEKPMFEIVCAKQTVIPNSEEVIHLHLRESGLMFYMTKDS
                     AATISNNIEACLVDVFKSVGMTPPEDWNSLFWIPHPGGRAILDQVEAKLKLRPEKFSAT
                     RTVLWDYGNMISACVLYILDEMRRKSAAEGLETYGEGLEWGVLLGFGPGMTIETILLHS
                     LPPV"
     gene            71..1282
                     /gene="PKS3"
                     /label=PKS3
     CDS             1293..1310
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1424..1442
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1443..1467
                     /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)."
     CDS             1576..1620
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      1672..1719
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      1673..1719
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     CDS             complement(1890..2681)
                     /codon_start=1
                     /product="spectinomycin resistance protein"
                     /label=spectinomycin resistance protein
                     /protein_id="AXN70005.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2682..2773)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2821..3559)
                     /direction=LEFT
                     /label=CDF
                     /note="CDF"
     CDS             complement(3769..4851)
                     /codon_start=1
                     /gene="lacIq"
                     /product="lac repressor protein"
                     /label=lacIq
                     /protein_id="AXN70006.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3769..4851)
                     /gene="lacIq"
                     /label=lacIq
     promoter        complement(4852..4929)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4975..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4975..4991
                     /regulatory_class="promoter"
                     /note="T7 promoter"