LOCUS       sequence-424944-        3276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PrpsM driven expression of ssrA-tagged eGFP (LVA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3276)
  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 3276)
  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: pEM7063
FEATURES             Location/Qualifiers
     source          1..3276
                     /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"
     CDS             1198..1230
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      1248..1334
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1371..1388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1584..1603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2087..2181)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2211..3002,3003..3071))
                     /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     2834..2853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3072..3176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3244..3262
                     /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 aaggcctgct
     1201 gcaaacgacg aaaactacgc tttagtagct taataatcta gaggcatcaa ataaaacgaa
     1261 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     1321 tgagtaggac aaatccgccg ccctagacct aggcggagat attccgcttc ctcgctcact
     1381 gactcgctcc gctcggtcgt tcggctgcgg cgagcggtac cggcttactg gcggggcgga
     1441 aatttcctgg aagatgccag gaagaaactt aacagggaag cgataaggcc acggcagagc
     1501 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
     1561 ggtggcgaaa cccgacagga cttaaagata ccaggcgttt ccccctggtg gctccctcgt
     1621 gcgctctcct gttcctgcct ttcggtttac cggtgtcact ccgctgttat ggccgcgttt
     1681 atctcattcc acgcctgaca ctcggttccg ggaaggcagt tcgctccaag ctggactgta
     1741 tgcacgaacc ccccgttcag tccgactgct gcgcctgttc cgataactat cgacttgagt
     1801 ccaagccgga aagacacgac aaagcgccac tggcagcagc cactggtaag agggtgtaca
     1861 agagattact gatgcagagt tcttgaagtc agagccaaac tacggctaaa ctggaaggac
     1921 gaatttggtg gcagcggtct tgtacaccct gttaccacgg ttcagaagtt ccccaactta
     1981 ctgaaccttc gaaaaaccac ctccccaggt ggttttttcg ttttcagagc aagagattac
     2041 gacgcaacgg aaaggatctc aagaggttca tcttactagt gcttggattc tcaccaataa
     2101 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     2161 ctggatctat caacaggagt ccaagcgagc tcgtaaactt ggtctgacag ttaccaatgc
     2221 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2281 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2341 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2401 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2461 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2521 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2581 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2641 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2701 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2761 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2821 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2881 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2941 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3001 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3061 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3121 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3181 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     3241 aaaaataggc gtatcacgag gccctttcgt cttcac
//
