LOCUS       40924_20051        3029 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFH2191, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3029)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J."; 
            date: "2001"; volume: "20"; issue: "21"; pages: "5853-5862"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (04-DEC-2000) Department of Microbiology, Technical University of 
            Denmark, Anker Engelunds Vej, Kgs. Lyngby DK-2800, Denmark"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /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
                     /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             283..996
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIVADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMHELYK"
     promoter        1124..1228
                     /label=AmpR promoter
     CDS             1229..2086
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /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"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtcgactct agaggatccc cgggtaccgc atatgtctaa aggtgaagaa
      301 ctgtttacgg gtgttgtgcc gatcctggtg gaactggatg gtgatgtgaa cggtcataaa
      361 ttctctgtgt ctggtgaagg tgaaggtgat gcaacctacg gtaagctgac cctgaaattt
      421 atctgtacca cgggtaaact gccggttcca tggccgaccc tggtgaccac cttcgcgtat
      481 ggtctgcaat gctttgcgcg ctacccggat cacatgaaac gtcacgactt tttcaaatct
      541 gccatgccgg aaggttatgt acaggaacgt accatcttct tcaaagatga cggtaactac
      601 aaaacccgtg ctgaagtgaa atttgaaggt gataccctgg tgaaccgtat cgaactgaaa
      661 ggtattgatt ttaaagaaga tggtaacatc ctgggccata aactggaata caactacaac
      721 tctcataacg tatacatcgt ggcagacaaa cagaaaaacg gtatcaaagt gaacttcaaa
      781 atccgtcata acatcgaaga tggttctgtt caactggcag accattatca gcagaacacc
      841 ccaattggcg atggcccggt gctgctgccg gacaaccatt acctgtccac gcagtctgcc
      901 ctttcgaaag atccgaacga aaaacgtgac cacatggtgc tgctggaatt tgtgaccgct
      961 gccggcatca cgcatggcat gcatgagctg tataaataaa gcatgcactg ggtggcggct
     1021 gcctaataat gcatatgagc ctaggacgcg tagctgcacg gaccgaggcc tactagtcgg
     1081 ccgtacgggc ccgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1381 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1441 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1741 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1981 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2101 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2401 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2881 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2941 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3001 cagcgagtca gtgagcgagg aagcggaag
//
