LOCUS       sequence-424906-        3237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PrpsM driven expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3237)
  AUTHORS   Delgadillo-Guevara M, Halte M, Erhardt M, Popp PF
  TITLE     Fluorescent tools for the standardized work in Gram-negative 
            bacteria.
  JOURNAL   J Biol Eng. 2024 Apr 8;18(1):25. doi: 10.1186/s13036-024-00420-9.
  PUBMED    38589953
REFERENCE   2  (bases 1 to 3237)
  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: "J Biol 
            Eng."; date: "2024-04-8"; volume: "18(1):25. doi"; pages: " 
            10.1186/s13036-024-00420-9"
COMMENT     Sequence Label: pEM13825
FEATURES             Location/Qualifiers
     source          1..3237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(478..480,481..1191)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(520..541)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(781..800)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1128..1149
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1209..1295
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1332..1349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1545..1564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2048..2142)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2172..2963,2964..3032))
                     /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     2795..2814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3033..3137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3205..3223
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgaggatc ccgggtacct gcagctagcg tcgacgaaag gctacggccg ttaattggtc
       61 gcctgagaag ttacggagag taaaaatgaa agttcgtgct tccgtcaaga aattatgccg
      121 taactgcaaa atcgttaagc gtgatggtgt catccgtgtg atttgcagtg ccgagccgaa
      181 gcataaacag cgccaaggct gattttttcg catatttttc ttgcaaagtt gggttgagct
      241 ggctagatta gccagccaat cttttgtatg tctgtacgtt tccatttgag tatcctgaaa
      301 acgggctttt cagcatggta cgtacatatt aaatagtagg agtgcatagt ggcccgtata
      361 gcaggcatta acattcctga tcagaaacac gccgtgatcg cgttaacttc gatctacggt
      421 gtcggcaaga cccgttctaa agccatcctg gctggaattc ataaggagga aaaacatatg
      481 agcaaaggcg aagaactgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      541 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      601 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      661 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      721 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      781 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      841 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      901 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
      961 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1021 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1081 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1141 gagttcgtga ccgccgccgg gatcactctc ggcatggatg aactgtataa ataataatct
     1201 agaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1261 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc taggcggaga
     1321 tattccgctt cctcgctcac tgactcgctc cgctcggtcg ttcggctgcg gcgagcggta
     1381 ccggcttact ggcggggcgg aaatttcctg gaagatgcca ggaagaaact taacagggaa
     1441 gcgataaggc cacggcagag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     1501 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg acttaaagat accaggcgtt
     1561 tccccctggt ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcac
     1621 tccgctgtta tggccgcgtt tatctcattc cacgcctgac actcggttcc gggaaggcag
     1681 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgactgc tgcgcctgtt
     1741 ccggtaacta tcgacttgag tccaagccgg aaagacacga caaagcgcca ctggcagcag
     1801 ccactggtaa gagggtgtac aagagattac tgatgcagag ttcttgaagt cagagccaaa
     1861 ctacggctaa actggaagga cgaatttggt ggcagcggtc ttgtacaccc tgttaccacg
     1921 gttcagaagt tccccaactt actgaacctt cgaaaaacca cctccccagg tggttttttc
     1981 gttttcagag caagagatta cgacgcaacg gaaaggatct caagaggttc atcttactag
     2041 tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     2101 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgtaaact
     2161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3181 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcac
//
