LOCUS       sequence-147419-        9292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for mf-lon mediated mCherry degradation in Lactococcus 
            lactis. Contains mcherry-pdt3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9292)
  AUTHORS   Cameron DE, Collins JJ
  TITLE     Tunable protein degradation in bacteria.
  JOURNAL   Nat Biotechnol. 2014 Dec;32(12):1276-81. doi: 10.1038/nbt.3053. Epub
            2014 Nov 17.
  PUBMED    25402616
REFERENCE   2  (bases 1 to 9292)
  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: "10.1038/nbt.3053"; 
            journalName: "Nat Biotechnol"; date: "2014-12"; volume: "32"; issue:
            "12"; pages: "1276-81"
COMMENT     Sequence Label: pECGMC3
FEATURES             Location/Qualifiers
     source          1..9292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2605..2632
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6152..6256
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6257..6325,6326..7117)
                     /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     complement(6475..6494)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7147..7241
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      7329..7917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7818..7837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(8082..8168)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctcgagagtc ttataactat actgacaata gaaacattaa caaatctaaa acagtcttaa
       61 ttctatcttg agaaagtatt ggtaataata ttattgtcga taacgcgagc ataataaacg
      121 gctctgatta aattctgaag tttgttagat acaatgattt cgttcgaagg aactacaaaa
      181 taccgcggtt aggggctagc atgagtaaaa aaataaaact acctattttc caaatacgcg
      241 gttcatttat tgttcctgga attaaagaga atttagaagt tggaagaaaa aacacattag
      301 cgagcgttaa ttatgcgatt aaaaattcaa ataatcaaat gatcgcaata ccgcagattg
      361 atgcatctgt tgaaaaacct gaattttcag atttacatga atttggaatt ttaattgatt
      421 ttgaagtaat caaagaatgg aaagataatt cattaactat tagcacaaac cctattcaac
      481 gatgtaaagt aattagcttt tttgaaaatg aagatcaagt gccgtatgcg gaagttgaat
      541 taattgaatc aattaatgat tttagtgatg aagagttaaa agaattaatt gaaaaaatct
      601 ctgatgctat caaaactaaa gctagtttag taacaaaaca aattaaacag ttaatatcag
      661 gtgaatctga tgatttaagt ttagcttttg attcaattat gtttaaactg gctccgtcaa
      721 aaatattgac aaatccggaa tacataacat ctccgtcatt aaaaacacga tggtcaatta
      781 ttgaaaaaat aatttttgct gaagatggaa tcataacaag aaatgcagaa tcaattgatg
      841 cagcacgaca aaagaatgaa attgaacaag aactgaacca taaattaaaa gaaaaaatgg
      901 ataaacaaca aaaagaatat tatctgcgag aaaaaatgcg aattatcaaa gatgaattag
      961 aagatgaaga tgattctgat gacagttcat tagaaaaata taaagaacgt ttagctaaag
     1021 aacctttccc tgaagaagtc aaacgaaaaa ttatggcttc tatcaaacga gttgaagcac
     1081 ttcaatcagg aactcctgaa tggaatactg agaaaaatta cattgattgg atgatgagta
     1141 ttccctggtg ggaagaaact gaagacttaa ctgatttaaa atatgctaaa aaaattttag
     1201 acaaacatca ttatggtatg aaaaaagtta aagaaagaat tattgagtac ttagctgtta
     1261 aaactaaaac aaaatcacta aaagcaccaa ttattacatt agttggtccc ccaggtgtag
     1321 gtaaaacaag tttagcaaaa tcaattgcag aagcagttgg aaaaaacttt gttaaagtaa
     1381 gtttaggtgg agttaaagat gaatcagaaa ttcgtggaca tagaaaaact tatgttggtt
     1441 caatgccagg aagaattatt caaacaatga aaagagcaaa agtaaaaaac ccattattct
     1501 tgcttgatga aattgataaa atggcatctg atcacagagg tgatccagca agtgcaatgt
     1561 tagaggttct tgacccagaa caaaacaaag agttttcaga tcattatatt gaagaaccat
     1621 atgatctaag ccaagttatg tttatagcaa ctgctaacta tcctgaagat attcctgaag
     1681 ccttatatga ccgtatggaa atcataaatt tatctagcta tactgaaatt gaaaaagtta
     1741 agatagcaca agattattta gttccaaaag caatagaaca acatgaatta acttcagaag
     1801 aaatatcatt tactgaaggt gcaattaatg aaattattaa atattacact agagaagctg
     1861 gtgtgcgtca attagaaaga catattaatt ctattattag aaagtatatt gttaaaaatc
     1921 taaatggtga aatggataaa attgttattg atgaaaaaca agtaaatgat ttattaggta
     1981 aaagaatttt tgatcacact gaaaaacaag aagaatctca aattggtgtg gttacaggtt
     2041 tagcatatac acaatttggt ggagatattt taccaattga agtaagttta taccctggta
     2101 aaggaaattt aattcttaca ggtaaacttg gtgaagttat gaaagaatct gcaacaattg
     2161 cgttaactta tgttaaatca aactttgaaa aatttggagt agataaaaaa gtatttgaag
     2221 aaaatgatat tcatgttcac gttcctgaag gagctgttcc aaaagatgga ccaagtgctg
     2281 gtatcacaat tacaactgct ttaatttctg cactttcaga taaaccagtt tcaaaagaaa
     2341 ttggaatgac tggtgaaatc acactaagag gaaatgtatt gccaattggt ggattaagag
     2401 aaaaatcaat ttctgcttca agaagtggat tgaaaacaat cattattcca aagaaaaatg
     2461 aaagagactt agacgaaatt ccagatgaag taaaagcaaa actaaaaatt atacctgctg
     2521 aaaaatatga agaggtattt gcaatagttt ttaaaacaaa ataagacgtc cctgagtagg
     2581 acaaatccgc cgctctagct aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     2641 tacaaactct tgttggggat ccgcctgcag cctgcggaaa cgtaaaagaa gttatggaaa
     2701 taagacttag aagcaaactt aagagtgtgt tgatagtgca gtatcttaaa attttgtata
     2761 ataggaattg aagttaaatt agatgctaaa aatttgtaat taagaaggag tgattacatg
     2821 aacaaaaata taaaatattc tcaaaacttt ttaacgagtg aaaaagtact caaccaaata
     2881 ataaaacaat tgaatttaaa agaaaccgat accgtttacg aaattggaac aggtaaaggg
     2941 catttaacga cgaaactggc taaaataagt aaacaggtaa cgtctattga attagacagt
     3001 catctattca acttatcgtc agaaaaatta aaactgaata ctcgtgtcac tttaattcac
     3061 caagatattc tacagtttca attccctaac aaacagaggt ataaaattgt tgggagtatt
     3121 ccttaccatt taagcacaca aattattaaa aaagtggttt ttgaaagcca tgcgtctgac
     3181 atctatctga ttgttgaaga aggattctac aagcgtacct tggatattca ccgaacacta
     3241 gggttgctct tgcacactca agtctcgatt cagcaattgc ttaagctgcc agcggaatgc
     3301 tttcatccta aaccaaaagt aaacagtgtc ttaataaaac ttacccgcca taccacagat
     3361 gttccagata aatattggaa gctatatacg tactttgttt caaaatgggt caatcgagaa
     3421 tatcgtcaac tgtttactaa aaatcagttt catcaagcaa tgaaacacgc caaagtaaac
     3481 aatttaagta ccgttactta tgagcaagta ttgtctattt ttaatagtta tctattattt
     3541 aacgggagga aataattcta tgagtcaccg gtataatgca cgaaatcgtt gcttattttt
     3601 ttttaaaagc ggtatactag atataacgaa acaacgaact gaatagaaac gaaaaaagag
     3661 ccatgacaca tttataaaat gtttgacgac attttataaa tgcatagccc gataagattg
     3721 ccaaaccaac gcttatcagt tagtcagatg aactcttccc tcgtaagaag ttatttaatt
     3781 aactttgttt gaagacggta tataaccgta ctatcattat atagggaaat cagagagttt
     3841 tcaagtatct aagctactga atttaagaat tgttaagcaa tcaatcggaa atcgtttgat
     3901 tgcttttttt gtattcattt atagaaggtg gagtttgtat gaatcatgat gaatgtaaaa
     3961 cttatataaa aaatagttta ttggagataa gaaaattagc aaatatctat acactagaaa
     4021 cgtttaagaa agagttagaa aagagaaata tctacttaga aacaaaatca gataagtatt
     4081 tttcttcgga gggggaagat tatatatata agttaataga aaataacaaa ataatttatt
     4141 cgattagtgg aaaaaaattg acttataaag gaaaaaaatc tttttcaaaa catgcaatat
     4201 tgaaacagtt gaatgaaaaa gcaaaccaag ttaattaaac aacctatttt ataggattta
     4261 taggaaagga gaacagctga atgaatatcc cttttgttgt agaaactgtg cttcatgacg
     4321 gcttgttaaa gtacaaattt aaaaatagta aaattcgctc aatcactacc aagccaggta
     4381 aaagcaaagg ggctattttt gcgtatcgct caaaatcaag catgattggc ggtcgtggtg
     4441 ttgttctgac ttccgaggaa gcgattcaag aaaatcaaga tacatttaca cattggacac
     4501 ccaacgttta tcgttatgga acgtatgcag acgaaaaccg ttcatacacg aaaggacatt
     4561 ctgaaaacaa tttaagacaa atcaatacct tctttattga ttttgatatt cacacggcaa
     4621 aagaaactat ttcagcaagc gatattttaa caaccgctat tgatttaggt tttatgccta
     4681 ctatgattat caaatctgat aaaggttatc aagcatattt tgttttagaa acgccagtct
     4741 atgtgacttc aaaatcagaa tttaaatctg tcaaagcagc caaaataatt tcgcaaaata
     4801 tccgagaata ttttggaaag tctttgccag ttgatctaac gtgtaatcat tttggtattg
     4861 ctcgcatacc aagaacggac aatgtagaat tttttgatcc taattaccgt tattctttca
     4921 aagaatggca agattggtct ttcaaacaaa cagataataa gggctttact cgttcaagtc
     4981 taacggtttt aagcggtaca gaaggcaaaa aacaagtaga tgaaccctgg tttaatctct
     5041 tattgcacga aacgaaattt tcaggagaaa agggtttaat agggcgtaat aacgtcatgt
     5101 ttaccctctc tttagcctac tttagttcag gctattcaat cgaaacgtgc gaatataata
     5161 tgtttgagtt taataatcga ttagatcaac ccttagaaga aaaagaagta atcaaaattg
     5221 ttagaagtgc ctattcagaa aactatcaag gggctaatag ggaatacatt accattcttt
     5281 gcaaagcttg ggtatcaagt gatttaacca gtaaagattt atttgtccgt caagggtggt
     5341 ttaaattcaa gaaaaaaaga agcgaacgtc aacgtgttca tttgtcagaa tggaaagaag
     5401 atttaatggc ttatattagc gaaaaaagcg atgtatacaa gccttattta gtgacgacca
     5461 aaaaagagat tagagaagtg ctaggcattc ctgaacggac attagataaa ttgctgaagg
     5521 tactgaaggc gaatcaggaa attttcttta agattaaacc aggaagaaat ggtggcattc
     5581 aacttgctag tgttaaatca ttgttgctat cgatcattaa agtaaaaaaa gaagaaaaag
     5641 aaagctatat aaaggcgctg acaaattctt ttgacttaga gcatacattc attcaagaga
     5701 ctttaaacaa gctagcagaa cgccctaaaa cggacacaca actcgatttg tttagctatg
     5761 atacaggctg aaaataaaac ccgcactatg ccattacatt tatatctatg atacgtgttt
     5821 gttttttctt tgctgtttag cgaatgatta gcagaaatat acagagtaag attttaatta
     5881 attattaggg ggagaaggag agagtagccc gaaaactttt agttggcttg gactgaacga
     5941 agtgagggaa aggctactaa aacgtcgagg ggcagtgaga gcgaagcgaa cacttgattt
     6001 tttaattttc tatcttttat aggtcattag agtatactta tttgtcctat aaactattta
     6061 gcagcataat agatttattg aataggtcat ttaagttgag catattagag gaggaaaatc
     6121 gcatgcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6181 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6241 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6301 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6361 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6421 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6481 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6541 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6601 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6661 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6721 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6781 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6841 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6901 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6961 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7021 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7081 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttacgagct
     7141 cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     7201 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact agtcatgacc
     7261 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7321 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7381 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7441 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     7501 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7561 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7621 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7681 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     7741 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7801 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7861 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7921 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7981 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8041 accgctcgcc gcagccgaac gccctaggtc tagggcggcg gatttgtcct actcaggaga
     8101 gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     8161 tttatttgat gcctctagca cgcgtggaac ctctagatta aactcgtctt ctgttcgcct
     8221 ggcccgcgtt cagcataaag gtcggcactt cgttggtgtt ttcttcgttt ttgttcgccg
     8281 ctttgtaaag ttcatccata ccaccagtag agtgacgacc ttcagcacgt tcgtattgtt
     8341 caacgatagt gtaatcttca ttgtgagaag tgatatcaag tttgatatta acattgtaag
     8401 caccaggaag ttgaacaggt tttttagctt tgtaagtagt tttaacttca gcatcgtagt
     8461 gaccaccatc tttaagttta agacgttgtt tgatttcacc tttaagagca ccatcttcag
     8521 ggtacatacg ttcagaagat gcttcccaac ccatagtttt tttttgcata acaggaccat
     8581 cagaagggaa attagtacca cgaagtttaa ctttgtagat aaattcacca tcttgaagag
     8641 aagaatcttg agtaacagta acaacaccac catcttcgaa attcataaca cgttcccatt
     8701 tgaaaccttc agggaaagaa agtttaaggt aatcagggat atcagcaggg tgtttaacgt
     8761 aggctttaga accgtacatg aattgaggag aaaggatatc ccaagcgaaa ggaagaggac
     8821 cacctttagt aactttaagt ttagcagttt gagtaccttc gtaaggacga ccttcacctt
     8881 caccttcgat ttcaaattcg tgaccattaa cagaaccttc catgtgaact ttgaaacgca
     8941 taaattcttt gatgatagcc atattatctt cttcaccttt agaaaccatg gtaccacctc
     9001 gctctttaat gaattcccga atattttttt acctacctag tatagcattt tgtgaagttt
     9061 ttttctagtc caagctcaca aaaatccaaa gtaaccgctt tattaagcca ttcttaaata
     9121 aaaataaaaa aagattaata gctaaaacta ttaatcttat catatcccga ggaccggttt
     9181 cgaaccggta cgaaggttac ccctcgcagg attttaagtc ctgtgcgtct gccagttccg
     9241 ccaccccggg tattatacaa aaactcccgt agggagattt atcaagcgaa cg
//
