LOCUS       sequence-160287-        4142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan-green fluorescent protein with tryptophan-based chromophore in 
            equilibrium between anionic and neutral states..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4142)
  AUTHORS   Sarkisyan KS, Yampolsky IV, Solntsev KM, Lukyanov SA, Lukyanov KA, 
            Mishin AS
  TITLE     Tryptophan-based chromophore in fluorescent proteins can be anionic.
  JOURNAL   Sci Rep. 2012;2:608. doi: 10.1038/srep00608. Epub 2012 Aug 29.
  PUBMED    22934131
REFERENCE   2  (bases 1 to 4142)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2012"; volume: "2:608. doi"; pages: " 10.1038/srep00608. Epub
            2012 Aug 29"
COMMENT     Sequence Label: WasCFP /pQE-30
FEATURES             Location/Qualifiers
     source          1..4142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..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             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(151..153,154..867)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLKTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISGNVNITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKQSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(193..214)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(454..473)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     801..822
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      889..983
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1027..1686
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1093..1112)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1751..1837
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1884..1906)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1992..2132
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2047..2066
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2147..2164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2318..2906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2398..2417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3077..3868,3869..3937))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3700..3719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3938..4042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4110..4128
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc gtgagcaagg gcgaggagct gttcaccggg
      181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      301 ggcaagctgc ccgtgccctg gcccaccctc aaaaccaccc tgacctgggg cgtgcagtgc
      361 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acgccatcag cggcaacgtc
      601 aatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
      661 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      721 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagca gagcaaagac
      781 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
      841 ctcggcatgg acgagctgta caagtaaaag cttaattagc tgagcttgga ctcctgttga
      901 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
      961 gggcgttttt tattggtgag aatccaagct agcttggcga gattttcagg agctaaggaa
     1021 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1081 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1141 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1201 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
     1261 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1321 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1381 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1441 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     1501 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1561 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1621 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     1681 gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggggtaat gactctctag
     1741 cttgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1801 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccctctagag ctgcctcgcg
     1861 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     1921 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1981 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     2041 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     2101 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     2161 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2221 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2281 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2341 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2401 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2461 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2521 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2581 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2641 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2701 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     2761 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2821 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2881 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2941 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3001 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3061 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3121 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3181 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3241 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3301 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3361 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3421 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3481 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3541 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3601 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3661 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3721 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3781 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3841 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3901 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3961 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4021 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4081 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     4141 ac
//
