LOCUS       sequence-424946-        3279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PrpsM driven expression of ssrA-tagged mCerulean (LVA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3279)
  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 3279)
  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: pEM7065
FEATURES             Location/Qualifiers
     source          1..3279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1201..1233
                     /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      1251..1337
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1374..1391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1587..1606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2090..2184)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2214..3005,3006..3074))
                     /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     2837..2856
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3075..3179)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3247..3265
                     /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 gttagtaaag gggaggagtt atttactggt gtagttccga ttttagtcga gcttgacggg
      541 gatgtaaatg ggcacaaatt tagtgtctct ggtgaaggcg agggtgatgc tacttacggg
      601 aaacttacgc tgaagtttat ctgtaccact ggaaagcttc cggttccctg gcccaccttg
      661 gttacaacat tgacatgggg tgtccaatgc ttcgcgcgct atcccgatca tatgaaacag
      721 cacgatttct tcaagtctgc catgccggag ggttacgttc aagaacgcac aatctttttc
      781 aaggatgatg gaaattacaa aactcgtgcg gaagtaaagt tcgagggtga tacccttgtg
      841 aaccgtattg agttgaaggg gattgatttt aaagaggacg gcaacatttt agggcacaaa
      901 ttagagtata atgctatctc ggacaacgta tatatcacag cggacaaaca gaaaaacggc
      961 atcaaagcta acttcaaaat tcgtcataat attgaggatg ggtctgttca actggcggat
     1021 cattaccagc agaacactcc aattggggac ggcccagtct tgttaccgga taaccattat
     1081 ttgagcacgc agtctaaatt gtcgaaagat cccaatgaaa aacgggatca catggtactg
     1141 ttggagttcg tcacggcggc cgggattact ttggggatgg acgagttata taagaggcct
     1201 gctgcaaacg acgaaaacta cgctttagta gcttaataat ctagaggcat caaataaaac
     1261 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1321 tcctgagtag gacaaatccg ccgccctaga cctaggcgga gatattccgc ttcctcgctc
     1381 actgactcgc tccgctcggt cgttcggctg cggcgagcgg taccggctta ctggcggggc
     1441 ggaaatttcc tggaagatgc caggaagaaa cttaacaggg aagcgataag gccacggcag
     1501 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     1561 agtggtggcg aaacccgaca ggacttaaag ataccaggcg tttccccctg gtggctccct
     1621 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc actccgctgt tatggccgcg
     1681 tttatctcat tccacgcctg acactcggtt ccgggaaggc agttcgctcc aagctggact
     1741 gtatgcacga accccccgtt cagtccgact gctgcgcctg ttccggtaac tatcgacttg
     1801 agtccaagcc ggaaagacac gacaaagcgc cactggcagc agccactggt aagagggtgt
     1861 acaagagatt actgatgcag agttcttgaa gtcagagcca aactacggct aaactggaag
     1921 gacgaatttg gtggcagcgg tcttgtacac cctgttacca cggttcagaa gttccccaac
     1981 ttactgaacc ttcgaaaaac cacctcccca ggtggttttt tcgttttcag agcaagagat
     2041 tacgacgcaa cggaaaggat ctcaagaggt tcatcttact agtgcttgga ttctcaccaa
     2101 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     2161 ttactggatc tatcaacagg agtccaagcg agctcgtaaa cttggtctga cagttaccaa
     2221 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2281 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2341 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2401 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2461 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2521 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2581 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2641 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2701 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2761 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2821 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2881 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2941 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3001 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3061 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3121 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3181 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3241 tataaaaata ggcgtatcac gaggcccttt cgtcttcac
//
