LOCUS       Exported                4955 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCDF-RpALS, complete sequence.
ACCESSION   MH488925
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4955)
  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 4955)
  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 4955)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4955
                     /organism="Cloning vector pCDF-RpALS"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:2301142"
     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..1246
                     /codon_start=1
                     /gene="ALS"
                     /product="aloesone synthase"
                     /label=ALS
                     /note="derived from Rheum palmatum"
                     /protein_id="AXN70001.1"
                     /translation="MADVLQEIRNSQKASGPATVLAIGTAHPPTCYPQADYPDFYFRVC
                     KSEHMTKLKKKMQFICDRSGIRQRFMFHTEENLGKNPGMCTFDGPSLNARQDMLIMEVP
                     KLGAEAAEKAIKEWGQDKSRITHLIFCTTTSNDMPGADYQFATLFGLNPGVSRTMVYQQ
                     GCFAGGTVLRLVKDIAENNKGARVLVVCSEIVAFAFRGPHEDHIDSLIGQLLFGDGAAA
                     LVVGTDIDESVERPIFQIMSATQATIPNSLHTMALHLTEAGLTFHLSKEVPKVVSDNME
                     ELMLEAFKPLGITDWNSIFWQVHPGGRAILDKIEEKLELTKDKMRDSRYILSEYGNLTS
                     ACVLFVMDEMRKRSFREGKQTTGDGYEWGVAIGLGPGLTVETVVLRSVPIP"
     gene            71..1246
                     /gene="ALS"
                     /label=ALS
     CDS             1257..1274
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1388..1406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1407..1431
                     /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             1540..1584
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      1636..1683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      1637..1683
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     CDS             complement(1854..2645)
                     /codon_start=1
                     /product="spectinomycin resistance protein"
                     /label=spectinomycin resistance protein
                     /protein_id="AXN70002.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2646..2737)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2785..3523)
                     /direction=LEFT
                     /label=CDF
                     /note="CDF"
     CDS             complement(3733..4815)
                     /codon_start=1
                     /gene="lacIq"
                     /product="lac repressor protein"
                     /label=lacIq
                     /protein_id="AXN70003.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3733..4815)
                     /gene="lacIq"
                     /label=lacIq
     promoter        complement(4816..4893)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4939..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4939..4955
                     /regulatory_class="promoter"
                     /note="T7 promoter"