LOCUS       Exported                4694 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pFHC2101, complete sequence.
ACCESSION   AY070364
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4694)
  AUTHORS   Atlung T, Hansen FG.
  TITLE     Effect of different concentrations of H-NS protein on chromosome 
            replication and the cell cycle in Escherichia coli
  JOURNAL   J. Bacteriol. 184 (7), 1843-1850 (2002)
  PUBMED    11889089
REFERENCE   2  (bases 1 to 4694)
  AUTHORS   Hansen FG, Atlung T.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2001) BioCentrum-DTU, Technical University of 
            Denmark, Building 301, Lyngby DK-2800, Denmark
REFERENCE   3  (bases 1 to 4694)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4694
                     /organism="Expression vector pFHC2101"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:183711"
     misc_feature    1
                     /label=SP6 transcription initiation site
                     /note="SP6 transcription initiation site"
     misc_feature    10..74
                     /note="multiple cloning site; synthetic adapters"
     gene            58..1543
                     /gene="lacI"
                     /label=lacI
     regulatory      58..64
                     /regulatory_class="ribosome_binding_site"
                     /gene="lacI"
                     /note="Shine-Dalgarno sequence"
     misc_feature    89..107
                     /gene="lacI"
                     /label=lac operator (O)
                     /note="lac operator (O)"
     protein_bind    91..107
                     /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      complement(116..144)
                     /regulatory_class="minus_10_signal"
                     /note="lacP(A1/04/03)"
     misc_feature    121..139
                     /gene="lacI"
                     /label=lac operator (O)
                     /note="lac operator (O)"
     protein_bind    123..139
                     /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)."
     promoter        383..460
                     /gene="lacI"
                     /label=lacI promoter
     CDS             461..1543
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="AAL59002.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTSKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     rep_origin      complement(1765..2540)
                     /direction=LEFT
     protein_bind    1850..1858
                     /label=DnaA binding site
                     /bound_moiety="DnaA"
     rep_origin      complement(1945..2533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1946
                     /label=RNA-DNA transition site
                     /note="RNA-DNA transition site"
     CDS             complement(2704..3564)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAL59003.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2704..3564)
                     /gene="bla"
                     /label=bla
     promoter        complement(3565..3669)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3949..3950
                     /note="inserted with Klenow polymerase to remove NdeI site"
     misc_feature    4005..4460
                     /label=phage f1 region
                     /note="phage f1 region"
     rep_origin      4005..4460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     regulatory      4540..4630
                     /regulatory_class="terminator"
                     /note="T7 terminator region"
     terminator      4559..4606
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        4678..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      4678..4694
                     /regulatory_class="promoter"
                     /note="SP6 promoter"