LOCUS       sequence-424922-        3270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PrpsM driven expression of ssrA-tagged mNeonGreen (LVA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3270)
  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 3270)
  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: pEM7064
FEATURES             Location/Qualifiers
     source          1..3270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             483..494
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1192..1224
                     /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      1242..1328
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1365..1382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1578..1597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2081..2175)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2205..2996,2997..3065))
                     /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     2828..2847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3066..3170)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3238..3256
                     /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 gtatcgaagg gcgaggaaga taacatggcg tcgttgccgg cgacccacga actgcatatt
      541 tttggcagta ttaacggcgt agactttgac atggtaggtc aggggacggg taatcctaat
      601 gatggctatg aagagttaaa cttaaagtcc acgaaagggg acctgcagtt cagcccttgg
      661 attctggtcc cacacattgg ctatggcttc caccagtact tgccttatcc ggacggcatg
      721 agtcccttcc aggccgcaat ggtcgatgga tcgggctacc aggtacatcg cacgatgcaa
      781 tttgaagacg gcgcaagtct gacggtgaac tatcgttata catatgaagg tagccatatt
      841 aaaggggaag cccaagtgaa agggactgga tttccggcag atggtccggt tatgactaat
      901 tcgctgactg cggcggattg gtgtcgttcc aaaaagacgt accccaatga caaaaccatc
      961 attagcactt ttaaatggag ctacaccaca ggcaacggga agcgctaccg cagtacggcc
     1021 cgtacgacgt atacttttgc caaaccgatg gcggcgaact acctgaagaa tcaaccgatg
     1081 tatgtctttc ggaaaacaga actgaagcac agtaagaccg agttaaactt caaagaatgg
     1141 caaaaagcat ttaccgatgt aatgggcatg gatgaactgt ataaaaggcc tgctgcaaac
     1201 gacgaaaact acgctttagt agcttaataa tctagaggca tcaaataaaa cgaaaggctc
     1261 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     1321 ggacaaatcc gccgccctag acctaggcgg agatattccg cttcctcgct cactgactcg
     1381 ctccgctcgg tcgttcggct gcggcgagcg gtaccggctt actggcgggg cggaaatttc
     1441 ctggaagatg ccaggaagaa acttaacagg gaagcgataa ggccacggca gagccgtttt
     1501 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     1561 gaaacccgac aggacttaaa gataccaggc gtttccccct ggtggctccc tcgtgcgctc
     1621 tcctgttcct gcctttcggt ttaccggtgt cactccgctg ttatggccgc gtttatctca
     1681 ttccacgcct gacactcggt tccgggaagg cagttcgctc caagctggac tgtatgcacg
     1741 aaccccccgt tcagtccgac tgctgcgcct gttccggtaa ctatcgactt gagtccaagc
     1801 cggaaagaca cgacaaagcg ccactggcag cagccactgg taagagggtg tacaagagat
     1861 tactgatgca gagttcttga agtcagagcc aaactacggc taaactggaa ggacgaattt
     1921 ggtggcagcg gtcttgtaca ccctgttacc acggttcaga agttccccaa cttactgaac
     1981 cttcgaaaaa ccacctcccc aggtggtttt ttcgttttca gagcaagaga ttacgacgca
     2041 acggaaagga tctcaagagg ttcatcttac tagtgcttgg attctcacca ataaaaaacg
     2101 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
     2161 ctatcaacag gagtccaagc gagctcgtaa acttggtctg acagttacca atgcttaatc
     2221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2341 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2521 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2821 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3001 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3061 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3181 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     3241 aggcgtatca cgaggccctt tcgtcttcac
//
