LOCUS       Exported                3029 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFH2191, complete sequence.
ACCESSION   AF325903
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3029)
  AUTHORS   Ostergaard H, Henriksen A, Hansen FG, Winther JR.
  TITLE     Shedding light on disulfide bond formation: engineering a redox 
            switch in green fluorescent protein
  JOURNAL   EMBO J. 20 (21), 5853-5862 (2001)
  PUBMED    11689426
REFERENCE   2  (bases 1 to 3029)
  AUTHORS   Hansen FG.
  TITLE     Synthetic Escherichia coli codon-optimized green fluorescent protein
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3029)
  AUTHORS   Hansen FG.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2000) Department of Microbiology, Technical 
            University of Denmark, Anker Engelunds Vej, Kgs. Lyngby DK-2800, 
            Denmark
REFERENCE   4  (bases 1 to 3029)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3029
                     /organism="Cloning vector pFH2191"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:153040"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             217..999
                     /codon_start=1
                     /product="lacZ-GFP fusion protein"
                     /label=lacZ-GFP fusion protein
                     /note="green fluorescent protein gene codons optimized for 
                     Escherichia coli"
                     /protein_id="AAK13447.1"
                     /translation="MTMITPSLHACRSTLEDPRVPHMSKGEELFTGVVPILVELDGDVN
                     GHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHD
                     FFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKL
                     EYNYNSHNVYIVADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY
                     LSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMHELYK"
     misc_feature    217..282
                     /label=Region: lacZ
                     /note="Region: lacZ"
     promoter        1124..1228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1229..2089
                     /codon_start=1
                     /product="beta-lactamase"
                     /function="confers resistance to ampicillin"
                     /label=beta-lactamase
                     /protein_id="AAK13448.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2260..2848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"