LOCUS       Exported                3776 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNCO113-hisactophilin, complete sequence.
ACCESSION   AF544403
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3776)
  AUTHORS   Kay CW, Schleicher E, Kuppig A, Hofner H, Rudiger W, Schleicher M, 
            Fischer M, Bacher A, Weber S, Richter G.
  TITLE     Blue light perception in plants. Detection and characterization of a
            light-induced neutral flavin radical in a C450A mutant of 
            phototropin
  JOURNAL   J. Biol. Chem. 278 (13), 10973-10982 (2003)
  PUBMED    12525505
REFERENCE   2  (bases 1 to 3776)
  AUTHORS   Fischer MJ, Bacher A.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-SEP-2002) Organic Chemistry and Biochemistry, 
            Technical University Munich, Lichtenbergstr. 4, Garching 85747, 
            Germany
REFERENCE   3  (bases 1 to 3776)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          join(1..114,499..3776)
                     /organism="Expression vector pNCO113"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:167123"
     source          1..3776
                     /organism="Expression vector pNCO113-hisactophilin"
                     /focus
                     /mol_type="genomic DNA"
                     /db_xref="taxon:209238"
     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..498
                     /codon_start=1
                     /product="hisactophilin"
                     /label=hisactophilin
                     /note="histidine rich protein from Dictyostelium 
                     discoideum; amino terminus fusions with product of interest
                     allows purfication with Ni-NTA"
                     /protein_id="AAN59911.1"
                     /translation="MGNRAFKSHHGHFLSAEGEAVKTHHGHHDHHTHFHVENHGGKVAL
                     KTHCGKYLSIGDHKQVYLSHHLHGDHSLFHLEHHGGKVSIKGHHHHYISADHHGHVSTK
                     EHHDHDTTFEEIIIGSAAASSGS"
     terminator      518..612
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1384..1470
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1626..1766
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1952..2540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2711..3571)
                     /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(3572..3676)
                     /gene="bla"
                     /label=AmpR promoter