LOCUS       Exported                3792 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNCO113-hisactophilin-C49S, complete sequence.
ACCESSION   EF469737
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3792)
  AUTHORS   Eisenreich W, Joshi M, Illarionov B, Richter G, Romisch-Margl W, 
            Muller F, Bacher A, Fischer M.
  TITLE     (13)C Isotopologue editing of FMN bound to phototropin domains
  JOURNAL   FEBS J. 274 (22), 5876-5890 (2007)
  PUBMED    17944933
REFERENCE   2  (bases 1 to 3792)
  AUTHORS   Fischer M.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-MAR-2007) Chemistry, University of Hamburg, 
            Grindelallee 117, Hamburg 20146, Germany
REFERENCE   3  (bases 1 to 3792)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3792
                     /organism="Expression vector pNCO113-hisactophilin-C49S"
                     /focus
                     /mol_type="other DNA"
                     /db_xref="taxon:452088"
     source          1..114
                     /organism="Expression vector pNCO113"
                     /mol_type="other DNA"
                     /db_xref="taxon:167123"
     source          513..3792
                     /organism="Expression vector pNCO113"
                     /mol_type="other DNA"
                     /db_xref="taxon:167123"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     protein_bind    62..78
                     /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)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             115..468
                     /codon_start=1
                     /product="hisactophilin C49S mutant"
                     /label=hisactophilin C49S mutant
                     /note="from Dictyostelium discoideum"
                     /protein_id="ABR45899.1"
                     /translation="MGNRAFKSHHGHFLSAEGEAVKTHHGHHDHHTHFHVENHGGKVAL
                     KTHSGKYLSIGDHKQVYLSHHLHGDHSLFHLEHHGGKVSIKGHHHHYISADHHGHVSTK
                     EHHDHDTTFEEIII"
     misc_feature    469..512
                     /label=linker
                     /note="linker"
     CDS             487..504
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      534..628
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1400..1486
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1642..1782
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1968..2556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2727..3587)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3588..3692)
                     /gene="bla"
                     /label=AmpR promoter