LOCUS       sequence-161782-        4110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NowGFP gene for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4110)
  AUTHORS   Sarkisyan KS, Goryashchenko AS, Lidsky PV, Gorbachev DA, Bozhanova 
            NG, Gorokhovatsky AY, Pereverzeva AR, Ryumina AP, Zherdeva VV, 
            Savitsky AP, Solntsev KM, Bommarius AS, Sharonov GV, Lindquist JR, 
            Drobizhev M, Hughes TE, Rebane A, Lukyanov KA, Mishin AS
  TITLE     Green fluorescent protein with anionic tryptophan-based chromophore 
            and long fluorescence lifetime.
  JOURNAL   Biophys J. 2015 Jul 21;109(2):380-9. doi: 10.1016/j.bpj.2015.06.018.
  PUBMED    26200874
REFERENCE   2  (bases 1 to 4110)
  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"; doi: 
            "10.1016/j.bpj.2015.06"; journalName: "Biophys J"; date: 
            "2015-07-21- 21"; volume: "109"; issue: "2"; pages: "380-9"
COMMENT     Sequence Label: NowGFP /pQE-30
FEATURES             Location/Qualifiers
     source          1..4110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..49
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    93..109
                     /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             128..139
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(182..184,185..898)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEKLFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKMSLK
                     FICTTGKLPVPWPTLKTTLTWGMQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNAISGNANITADKQKNGIKA
                     YFTIRHDVEDGSVLLADHYQQNTPIGDGPVLLPDNHYLSTQSKQSKDPNEKRDHMVLLE
                     FVTAAGIPLGADELYK"
     primer_bind     complement(224..245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(485..504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     832..853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      920..1014
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1058..1717
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1124..1143)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1782..1868
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1915..1937)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2023..2163
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2078..2097
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2178..2195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2349..2937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2429..2448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3108..3899,3900..3968))
                     /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     3731..3750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3969..4073)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
       61 cacctcgaga aatcataaaa aatttatttg ctttgtgagc ggataacaat ttcacacaga
      121 attcattaaa gaggagaaat taactatgag aggatcgcat caccagcacc atcacggatc
      181 cgtgagcaag ggcgagaagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      241 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      301 caagatgtcc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      361 caaaaccacc ctgacctggg gcatgcagtg cttcgcccgc taccccgacc acatgaagca
      421 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      481 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      541 gaaccgcatc gagctgaagg gcgtcgactt caaggaggac ggcaacatcc tggggcacaa
      601 gctggagtac aacgccatca gcggcaacgc caatatcacc gccgacaagc agaagaacgg
      661 catcaaggcc tacttcacga tccgccacga cgtcgaggac ggcagcgtgc tgctcgccga
      721 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
      781 cctgagcacc cagtccaagc agagcaaaga ccccaacgag aagcgcgatc acatggtcct
      841 gctggagttc gtgaccgccg ccgggatccc tctcggcgcg gacgagctgt acaagtaaaa
      901 gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat
      961 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
     1021 tagcttggcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     1081 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     1141 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     1201 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     1261 tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     1321 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     1381 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     1441 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     1501 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     1561 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     1621 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     1681 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
     1741 cccttaaacg cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc
     1801 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     1861 ggacaaatcc gccctctaga gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     1921 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1981 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     2041 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
     2101 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     2161 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2761 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     2821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3241 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4081 ccccgaaaag tgccacctga cgtctaagaa
//
