LOCUS       sequence-424605-        3258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PrpsM driven expression of ssrA-tagged mScarlet-I (LVA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3258)
  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 3258)
  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: pEM7066
FEATURES             Location/Qualifiers
     source          1..3258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1180..1212
                     /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      1230..1316
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1353..1370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1566..1585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2069..2163)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2193..2984,2985..3053))
                     /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     2816..2835
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3054..3158)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3226..3244
                     /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 gtgagcaaag gcgaagcggt gattaaagaa tttatgcgct ttaaagtgca tatggaaggc
      541 agcatgaacg gccatgaatt tgaaattgaa ggcgaaggcg aaggccgccc gtatgaaggc
      601 acccagaccg cgaaactgaa agtgaccaaa ggcggcccgc tgccgtttag ctgggatatt
      661 ctgagcccgc agtttatgta tggcagccgc gcgtttatta aacatccggc ggatattccg
      721 gattattata aacagagctt tccggaaggc tttaaatggg aacgcgtgat gaactttgaa
      781 gatggcggcg cggtgaccgt gacccaggat accagcctgg aagatggcac cctgatttat
      841 aaagtgaaac tgcgcggcac caactttccg ccggatggcc cggtgatgca gaaaaaaacc
      901 atgggctggg aagcgagcac cgaacgcctg tatccggaag atggcgtgct gaaaggcgat
      961 attaaaatgg cgctgcgcct gaaagatggc ggccgctatc tggcggattt taaaaccacc
     1021 tataaagcga aaaaaccggt gcagatgccg ggcgcgtata acgtggatcg caaactggat
     1081 attaccagcc ataacgaaga ttataccgtg gtggaacagt atgaacgcag cgaaggccgc
     1141 catagcaccg gcggcatgga tgaactgtat aaaaggcctg ctgcaaacga cgaaaactac
     1201 gctttagtag cttaataatc tagaggcatc aaataaaacg aaaggctcag tcgaaagact
     1261 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1321 cgccctagac ctaggcggag atattccgct tcctcgctca ctgactcgct ccgctcggtc
     1381 gttcggctgc ggcgagcggt accggcttac tggcggggcg gaaatttcct ggaagatgcc
     1441 aggaagaaac ttaacaggga agcgataagg ccacggcaga gccgtttttc cataggctcc
     1501 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     1561 gacttaaaga taccaggcgt ttccccctgg tggctccctc gtgcgctctc ctgttcctgc
     1621 ctttcggttt accggtgtca ctccgctgtt atggccgcgt ttatctcatt ccacgcctga
     1681 cactcggttc cgggaaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc
     1741 agtccgactg ctgcgcctgt tccggtaact atcgacttga gtccaagccg gaaagacacg
     1801 acaaagcgcc actggcagca gccactggta agagggtgta caagagatta ctgatgcaga
     1861 gttcttgaag tcagagccaa actacggcta aactggaagg acgaatttgg tggcagcggt
     1921 cttgtacacc ctgttaccac ggttcagaag ttccccaact tactgaacct tcgaaaaacc
     1981 acctccccag gtggtttttt cgttttcaga gcaagagatt acgacgcaac ggaaaggatc
     2041 tcaagaggtt catcttacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
     2101 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     2161 gtccaagcga gctcgtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2221 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2281 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2341 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2401 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2461 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2521 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2581 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2641 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2701 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2761 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2821 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2881 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2941 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3001 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3061 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3121 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3181 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3241 aggccctttc gtcttcac
//
