LOCUS       sequence-326670-        4118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a monomeric enhanced GFP variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4118)
  AUTHORS   Valbuena FM, Fizgerald I, Strack RL, Andruska N, Smith L, Glick BS
  TITLE     A photostable monomeric superfolder GFP.
  JOURNAL   Traffic. 2020 May 15. doi: 10.1111/tra.12737.
  PUBMED    32415747
REFERENCE   2  (bases 1 to 4118)
  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: "Traffic. 
            2020 May 15. doi: 10.1111/tra.12737."
COMMENT     Sequence Label: pQE-60NA-mEGFP
FEATURES             Location/Qualifiers
     source          1..4118
                     /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             join(115..117,118..120,121..834)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(160..181)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(421..440)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     768..789
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      865..959
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1069..1088)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1731..1817
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1864..1886)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1972..2112
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2027..2046
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2127..2144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2298..2886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2378..2397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3057..3848,3849..3917))
                     /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     3680..3699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3918..4022)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4086..4104
                     /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 aaccatggtg
      121 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      181 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      241 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      301 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      361 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      421 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      481 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      541 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
      601 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
      661 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      721 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
      781 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
      841 cgcaagctta attagctgag cttggactcc tgttgataga tccagtaatg acctcagaac
      901 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
      961 caagctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     1021 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     1081 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     1141 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     1201 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat
     1261 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     1321 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     1381 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     1441 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     1501 ttcgcccccg ttttcaccat gcatgggcaa atattatacg caaggcgaca aggtgctgat
     1561 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     1621 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     1681 ttattggtgc ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac
     1741 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1801 tcctgagtag gacaaatccg ccctctagag ctgcctcgcg cgtttcggtg atgacggtga
     1861 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     1921 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     1981 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     2041 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2101 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2161 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2221 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2281 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2701 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     2761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2881 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3121 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3181 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3241 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3301 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3361 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3421 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3481 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3541 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3601 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3661 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3721 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3781 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3841 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3901 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3961 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4021 cgcacatttc cccgaaaagt gccacctgct aagaaaccat tattatcatg acattaacct
     4081 ataaaaatag gcgtatcacg aggccctttc gtcttcac
//
