LOCUS       Exported                5898 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pET-hsp12, complete sequence.
ACCESSION   KY926444
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  AUTHORS   Leger A, Hocquellet A, Dieryck W, Moine V, Marchal A, Marullo P, 
            Josseaume A, Cabanne C.
  TITLE     Production and purification of the native Saccharomyces cerevisiae 
            Hsp12 in Escherichia coli
  JOURNAL   J. Agric. Food Chem. (2017) In press
  PUBMED    28871789
REFERENCE   2  (bases 1 to 5898)
  AUTHORS   Leger A, Hocquelet A, Dieryck W, Moine V, Marchal A, Josseaume A, 
            Cabanne C.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-APR-2017) ENSTBB, Institut Polytechnique de Bordeaux, 
            146 rue Leo Saignat, Bordeaux, Gironde 33076, France
REFERENCE   3  (bases 1 to 5898)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5898
                     /organism="Cloning vector pET-hsp12"
                     /mol_type="other DNA"
                     /note="yeast hsp12 with E. coli adaptated codons cloned 
                     into pET44a(+) vector"
                     /db_xref="taxon:2035513"
     misc_feature    1..4960
                     /label=pET-44a(+)
                     /note="pET-44a(+)"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      13..460
                     /note="/function=f1 origin"
     CDS             complement(582..1442)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicilin resistance protein"
                     /protein_id="ASZ84915.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1443..1535)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1594..2222
                     /label=ColE1
                     /note="ColE1"
     rep_origin      1616..2204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2390..2532
                     /label=bom
                     /note="basis of mobility region from pBR322"
     regulatory      2634..2825
                     /regulatory_class="replication_regulatory_region"
                     /note="rop"
     CDS             complement(2634..2825)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3397..4479)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             complement(3397..4356)
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /note="lactose operon repressor"
                     /protein_id="ASZ84916.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
     promoter        complement(4480..4557)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4870..4888
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4870..4886
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     protein_bind    4889..4913
                     /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)."
     regulatory      4891..4911
                     /regulatory_class="transcriptional_cis_regulatory_region"
                     /note="lacO"
     RBS             4928..4950
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      4945..4950
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Dalgarno"
     CDS             4958..5287
                     /codon_start=1
                     /product="yeast HSP12"
                     /label=yeast HSP12
                     /note="12 kDa heat shock protein; synthetic gene with 
                     Escherichia coli adapted codons"
                     /protein_id="ASZ84917.1"
                     /translation="MSDAGRKGFGEKASEALKPDSQKSYAEQGKEYITDKADKVAGKVQ
                     PEDNKGVFQGVHDSAEKGKDNAEGQGESLADQARDYMGAAKSKLNDAVEYVSGRVHGEE
                     DPTKK"
     misc_feature    5288..5898
                     /label=pET-44a(+)
                     /note="pET-44a(+)"
     CDS             5331..5363
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             5370..5387
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5553..5639
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      5556..5756
                     /regulatory_class="terminator"
                     /note="rrnB T1T2 terminator"
     terminator      5731..5758
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      5826..5873
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      5826..5873
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"