LOCUS       sequence-463667-        7982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lon protease from Mesoplasma florum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7982)
  AUTHORS   Sakkos JK, Hernandez-Ortiz S, Osteryoung KW, Ducat DC
  TITLE     Orthogonal Degron System for Controlled Protein Degradation in 
            Cyanobacteria.
  JOURNAL   ACS Synth Biol. 2021 Jul 16;10(7):1667-1681. doi: 
            10.1021/acssynbio.1c00035. Epub 2021 Jul 7.
  PUBMED    34232633
REFERENCE   2  (bases 1 to 7982)
  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"; doi: 
            "10.1021/acssynbio.1c00035"; journalName: "ACS Synth Biol"; date: 
            "2021-07-16- 16"; volume: "10"; issue: "7"; pages: "1667-1681"
COMMENT     Sequence Label: pEYN12-mflon
FEATURES             Location/Qualifiers
     source          1..7982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(301..306,307..323,324..330)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    338..354
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             439..504
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2941..2958)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3111..3197
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3289..3316
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4073..4090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4244..4832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4324..4343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5003..5794,5795..5863))
                     /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     5626..5645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5864..5968)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5994..6449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6131..6152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6343..6362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(7169..7982,1..2))
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLRFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7892..7911
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     join(7976..7982,1..13)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
       61 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata
      121 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
      181 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc
      241 aaccgtggct ccctcacttt ctggctggat gatggaattc aaatattctg aaatgagctg
      301 ttgacaatta atcatccggc tcgtataatg tgtggaattg tgagcggata acaatttcat
      361 acgctcacaa ttggtaccgg tgataccagc atcgtcttga tgcccttggc agcaccctgc
      421 taaggaggca acaagatgga ttataaagac cacgatggcg actataagga tcatgacatt
      481 gactacaaag atgatgatga taaaggtgcc ggcgctagca agaagattaa gctgcccatt
      541 tttcagattc gtggttcgtt tatcgtcccc ggcattaagg aaaacctcga agtcggccga
      601 aagaacaccc tggccagtgt caattatgct atcaaaaatt cgaataacca gatgatcgcc
      661 atcccacaaa tcgatgccag cgttgaaaaa cccgagttca gcgacctgca cgagttcggg
      721 attctcattg attttgaagt tatcaaggaa tggaaagaca attccctgac catttcgacc
      781 aaccccattc agcggtgcaa agtcatcagc ttctttgaga acgaagatca ggtgccatac
      841 gcggaggtcg agctcattga gagtatcaac gacttttctg atgaagaatt gaaagaactg
      901 attgaaaaaa ttagtgacgc catcaagact aaagcgtcgc tcgtgacgaa acaaattaag
      961 caattgatct cgggtgaaag tgacgatctg tcgttggcgt tcgattcgat catgtttaaa
     1021 ctggcgccgt ctaagatcct cactaatccc gagtatatta catctcccag tctcaagaca
     1081 cgctggtcga tcattgaaaa aattattttt gcggaagacg ggatcatcac ccggaatgct
     1141 gaatccattg acgcggcacg acaaaagaac gagattgaac aagagctgaa ccataaattg
     1201 aaagaaaaaa tggacaagca gcagaaggag tactatctgc gcgagaagat gcggattatc
     1261 aaggacgagt tggaagatga agatgactcc gatgacagtt ctctggagaa atataaggag
     1321 cgcttggcga aagagccctt ccccgaagag gtcaaacgaa agattatggc ttcgatcaag
     1381 cgcgtcgaag ccctgcaatc tggaaccccc gaatggaata ctgagaagaa ctacattgac
     1441 tggatgatga gtattccttg gtgggaagaa acggaggatc tgaccgattt gaagtacgct
     1501 aaaaagatcc tggacaagca tcattacgga atgaaaaaag tgaaagagcg aatcatcgaa
     1561 tatctcgctg tgaaaacgaa aaccaagagc ctgaaggccc ctatcattac actggtcggt
     1621 cctccgggcg ttgggaaaac tagcttggcg aaaagtatcg ctgaggcagt cggcaaaaat
     1681 tttgtcaagg tgagcttggg tggtgttaaa gacgagagcg aaatccgggg tcatcggaaa
     1741 acatatgtcg gttcgatgcc cggccgtatc atccaaacca tgaagcgggc caaagtgaaa
     1801 aacccattgt ttctcctcga tgaaatcgat aaaatggctt ctgatcatcg cggggaccca
     1861 gcatccgcca tgctcgaggt gctggatccc gagcaaaaca aagaattttc ggatcactat
     1921 attgaagaac cttacgatct ctctcaggtt atgttcattg cgacagcaaa ttacccagag
     1981 gatatccccg aagccctgta tgatcgcatg gagatcatta atctgtcgag ctacactgag
     2041 atcgagaagg tcaaaatcgc gcaagactat ctggtgccca aggcaattga acagcacgaa
     2101 ttgacgtctg aggaaatctc cttcacggaa ggcgctatta atgaaattat taagtactat
     2161 acccgggagg ccggcgtccg acaactcgaa cgtcatatca acagcattat tcggaaatat
     2221 attgtcaaaa atttgaacgg tgagatggat aagattgtga tcgacgaaaa gcaagttaat
     2281 gatttgctgg ggaaacgcat tttcgatcat actgaaaagc aggaggaatc tcaaattggc
     2341 gtggtcaccg gcctggccta cacgcagttt ggcggtgata ttttgcctat cgaagtgtct
     2401 ctgtaccccg gcaagggcaa tctcattttg acggggaaac tgggcgaggt gatgaaagaa
     2461 tcggcgacta ttgcgctgac ctatgttaaa tcgaacttcg aaaaattcgg tgtggacaaa
     2521 aaagtgtttg aagagaatga tatccatgtc cacgtccccg agggtgcagt gccaaaagac
     2581 ggccccagcg cgggaattac cattacaacc gccctcatca gcgcattgag tgataagcct
     2641 gtttccaaag agattggtat gacgggtgaa attacgctgc gaggaaacgt gctgcccatc
     2701 ggaggtttgc gggagaagtc gatttctgcc agccgtagcg gtctcaagac gattattatt
     2761 ccaaaaaaaa atgaacgcga tttggatgag attccggacg aggtcaaagc gaagctcaag
     2821 atcatccccg ccgaaaagta tgaagaggtg ttcgctattg tttttaagac caagtaattc
     2881 tagagtcgac ctgcaggcat gcaagcttgg ctgttttggc ggatgagaga agattttcag
     2941 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
     3001 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     3061 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     3121 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     3181 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     3241 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     3301 tgacggatgg cctttttgcg tttctacaaa ctcttgcggt tgctaccaag tagtgacccg
     3361 cttcgtgatg caaaatccgc tgacgatatt cgggcgatcg ctgctgaatg ccatcgagca
     3421 gtaacgtggc accccgcccc tgccaagtca ccgcatccag actgaacagc accaagaggc
     3481 taaaacccaa tcccgccggt agcagcggag aactacccag cattggtccc accaaagcta
     3541 atgccgtcgt ggtaaaaatc gcgatcgccg tcagactcaa gcccagttcg ctcatgcttc
     3601 ctcatctagg tcacagtctt cggcgatcgc atcgatctga tgctgcagca agcgttttcc
     3661 ataccggcga tcgcgccgtc gccctttcgc tgccgtggcc cgcttacgag ctcgtttatc
     3721 gaccacgatc gcatccaaat ccgcgatcgc ttcccagtcc ggcaattcag tctggggcgt
     3781 ccgtttcatt aatcctgatc aggcacgaaa ttgctgtgcg tagtatcgcg catagcggcc
     3841 agcctctgcc aacagcgcat cgtgattgcc tgcctcaaca atctggccgc gctccatcac
     3901 caagatgcgg ctggcattac gaaccgtagc cagacggtga gcaatgataa agaccgtccg
     3961 tccctgcatc acccgttcta ggacgtcttg caatcactag tgcggccctg cattaatgaa
     4021 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4081 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4141 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4201 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4261 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4321 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4381 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4441 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4501 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4561 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4621 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4681 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4741 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4801 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4861 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4921 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4981 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5041 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5101 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5161 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5221 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5281 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5341 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5401 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5461 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5521 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5581 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5641 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5701 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5761 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5821 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5881 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5941 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     6001 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6061 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6121 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6181 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6241 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6301 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6361 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6421 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     6481 ggaagggcga tcggtgcggg cctcttcgct attacgccag catggccgcg ggattctggc
     6541 ttacagggat aactgagagt caacagcctc tgtccgtcat tgcacaccca tccatgcact
     6601 ggggacttga ctcatgctga atcacatttc ccttgtccat tgggcgagag gggaggggaa
     6661 tcttctggac tcttcactaa gcggcgatcg caggttcttc tacccaagca gtggcgatcg
     6721 cttgattgca gtcttcaatg ctggcctctg cagccatcgc cgccaccaaa gcatcgtagg
     6781 cgggacgttg ttgctccagt aaagtcttcg cccgtaacaa tccccagcga ctgcgtaaat
     6841 ccgcttcggc aggattgcga tcgagttgcc gccacagttg tttccactgg gcgcgatcgt
     6901 cagctccccc ttccacgttg ccgtagacca gttgctctgc cgctgcaccg gccatcaaca
     6961 cctgacacca ctgttccagc gatcgctgac tgagttgccc ctgtgcggct tcggcttcta
     7021 gcgcagctgc ttggaactgc acacccccgc gaccaggttg tccttggcgc agcgcttccc
     7081 acgctgagag ggtgtagccc gtcacgggta actcccgtca agtcagcgta atgctctgcc
     7141 agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact
     7201 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     7261 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     7321 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     7381 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatcgcaaa agcttatgca
     7441 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     7501 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     7561 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     7621 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     7681 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     7741 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     7801 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     7861 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     7921 cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc
     7981 tc
//
