LOCUS       sequence-96226-e        7216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Disintegrator plasmid encoding MET17 promoter driven green beetle 
            luciferase with nuclear localization signal (NLS) and ClpXP degron 
            tag (ssrA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7216)
  AUTHORS   Mazo-Vargas A, Park H, Aydin M, Buchler NE
  TITLE     Measuring fast gene dynamics in single cells with time-lapse 
            luminescence microscopy
  JOURNAL   Mol Biol Cell. 2014 Sep 17. pii: mbc.E14-07-1187.
  PUBMED    25232010
REFERENCE   2  (bases 1 to 7216)
  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: "Mol Biol 
            Cell. 2014 Sep 17. pii: mbc.E14-07-1187."
COMMENT     Sequence Label: pNB695
FEATURES             Location/Qualifiers
     source          1..7216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..43)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(703..1494,1495..1563))
                     /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     1326..1345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1564..1668)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1736..1754
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1761..1981
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1982..2785
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(2903..2925)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3025..3044
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4084..4433
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             4455..4475
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6217..6404
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6272..6296)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6989..7006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7160..7216,1..532))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
       61 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      361 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
      421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1681 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     1741 gcgtatcacg aggccagctt ttcaattcaa ttcatcattt tttttttatt cttttttttg
     1801 atttcggttt ctttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag
     1861 gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa
     1921 ttgcccagta ttcttaaccc aactgcacag aacaaaaaca tgcaggaaac gaagataaat
     1981 catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa
     2041 gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac
     2101 caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac
     2161 acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt
     2221 atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac
     2281 agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa
     2341 tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt
     2401 aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc
     2461 tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat
     2521 cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat
     2581 gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt atagaaccgt
     2641 ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa
     2701 gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt
     2761 gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa
     2821 ctcacaaatt agagcttcaa tttaattata tcagttatta cccgcccttt cgtctcgcgc
     2881 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2941 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3001 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     3061 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     3121 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3181 aggcgtcgag aagtgacatc acccgaaaag aagctaagtc tttctcctaa ttcatattta
     3241 attattgtac atggacatat catacgtaat gctcaacctt aagctgctgc ggagcttcca
     3301 cgagcaccag cacctgaagc aactagactt atttgcgctt gagttagttc tatacttggc
     3361 agtggaagtt tgtcattatg agttggtgga ggtaaaaatc caaccttgtt taaaaatgca
     3421 atatatatac caacctcttc tggagtaaca cctattccat tagaccaatc aacacctgtc
     3481 catgcggtgt ctttctttaa agatgccacg gcgttcctat cgtctagttc cggagctttg
     3541 gtactttcag gtaagttgtc taagaccatg tgtagcaaag gatacaacgc attttcttca
     3601 ttcctgtcaa taacagacgc ctccaatgat ttcttccatt tagaatagct ttcgatttcg
     3661 acatcgtaac cataatcgtg taaagtatac aagtagtctt tgaataatat tcttgggtga
     3721 cccgttactt gagcaacggc caattcattt tctttgggag gattcaaaga cgtagcaaca
     3781 acaacacgag caacatgatc tactggaacc atattcacgg aattttcgat atctggaatc
     3841 ttacctaatt ggactgaacc tttcaaaaat ctcaataaga aatcatctgt gtttgaagaa
     3901 ccattggcag aggcacctgt tacgtaacct ggtctgacaa tacacccacg taggcccctt
     3961 tcacctgcac gtctaatgat gtactcagca gcccatttgg actgaccata tccaccagtg
     4021 agcccgcttg cagaggctgg aattcgagct ccggatgcaa gggttcgaat cccttagctc
     4081 tcattatttt ttgctttttc tcttgaggtc acatgatcgc aaaatggcaa atggcacgtg
     4141 aagctgtcga tattggggaa ctgtggtggt tggcaaatga ctaattaagt tagtcaaggc
     4201 gccatcctca tgaaaactgt gtaacataat aaccgaagtg tcgaaaaggt ggcaccttgt
     4261 ccaattgaac acgctcgatg aaaaaaataa gatatatata aggttaagta aagcgtctgt
     4321 tagaaaggaa gtttttcctt tttcttgctc tcttgtcttt tcatctacta tttccttcgt
     4381 gtaatacagg gtcgtcagat acatagatac aattctatta cccccatcca tactcttaat
     4441 taaccatggc tccaccaaaa aaaaaaagaa aagttgccat gggtgggggc agcatgatga
     4501 agagagaaaa aaatgttgtt tacggccccg aacccaaaca ccctttggga aactttactg
     4561 ctggagaaat gctctacaat gcccttcata agcattccca cataccgcaa gcaatattag
     4621 atgtgatggg taatgaatcg ctgtcatatc aagaattttt cgacactact gtcaagctag
     4681 gacaaagtct tcaaaattgt ggatacaaga tgaatgatgt agtgtcgatc tgtgctgaga
     4741 acaataaaag atttttcatc cccattattt cagcttggta tattggtatg gttgtagcac
     4801 ctgttaatga agactacatc ccagatgaac tttgtaaagt cacgggtata tcaaaaccaa
     4861 tactggtctt cactacaaga aaaatcttac ctaaggtatt agaggtaaag gacagaacta
     4921 attacataaa gagaattata atactagatt ctgaagaaaa tctgcttggt tgcgaaagtc
     4981 ttcataattt tatgtcccgt tattcggata ataatcttca gacctttaag cctctacatt
     5041 acgatcctgt tgatcaagtg gcagctattt tatgttcgtc aggcacaact ggattaccca
     5101 aaggtgttat gcaaacgcat agaaacattt gtgttcgact tacacatgct tcggatccca
     5161 gagttggaac acaacttatt cctggagtat cagttttggc gtatctgcct ttcttccatg
     5221 cttttgggtt ttctataaac ttaggatact ttatggtggg ccttcgtgtt gttatgctaa
     5281 gacgatttaa tcaagaagta tttttaaaag ccattcaaga ttatgaagtt cgaagtgtaa
     5341 tcaacgttcc atcaacaata ctgttcttgt cgaaaagtcc tttagttgac aaatacgatt
     5401 tatcgacttt ggcggaattg tgttgcggcg ctgcaccatt agcaaaggaa gttgctgaga
     5461 tagcagtgaa acgactaaac ctgccaggaa ttcgttgtgg atatggtttg acagagtcta
     5521 cttcagctaa tatacatact cttcacaatg aatttaagtc aggatcactt ggaaaagtca
     5581 ctccttatat ggctgcgaaa ataatagata ggaacactgg tgaagctttg ggaccaaatc
     5641 aagttggaga actatgcatc tggggtccta tggtaacaaa aggttacgtg aacaatccac
     5701 aagccaccaa agaggctatt gatgacgacg gttggcttca ctctggagac tttggatact
     5761 atgatgagga tgaatatttc tatatagtgg accgttacaa ggaacttatt aaatataaag
     5821 gctatcaggt agcacctgta gaattagaag agattttatt acaacatcca ggtataagag
     5881 atgttgctgt cgttggtatt cctgatatag aagctggaga actaccagct gggttcgtgg
     5941 ttaaacaacc cggagcacaa cttacagcaa aagaagttta cgattttctt gcccaacggg
     6001 tctctcattc aaagtatttg cgtggaggag ttcgattcgt tgattcaata cccaggaatg
     6061 ttacaggtaa aatttcaaga aaagaacttc gagaggcgtt gatggaaaaa gctggaggtg
     6121 gcagtatgta caaaactagt gcagcgaacg acgaaaatta cgcccttgca gcgtaataac
     6181 tagttaatac aaataaggcg cgccacttct aaataagcga atttcttatg atttatgatt
     6241 tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag
     6301 gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct
     6361 caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat
     6421 aacagggtaa tatagatctc tcgaggcggc cgccagcatc attgatgtaa tcttctacta
     6481 gttgatccaa ttgtccagca gctctaataa caatcaaccc acgtggttta gcaacaccta
     6541 agtaaatggt ttgtctggct gggggatatg cagggtcgat aactgaaaag gttgcgccgg
     6601 ctttcaagac acccatcaca cataccatca aatccacacc cctagaagaa tagatcatca
     6661 ctacatcacc tcttttgata cctgttttaa tcaaataatg ggcaactatg ttagaagtgc
     6721 ggttgatgtc gcgataagtg aaagaacggg acttgtcgga atttagtgtt ggagtctcca
     6781 caacacaggt tctctctggg aaggcttcag cattgtcctg gaaaatgtcg tgaatacacc
     6841 ccacgaaatc gcaccagccc aagttcttag ttggatcagg taaactatcc ttggatgatg
     6901 cggtgatcag agagaggatc tgcctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6961 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7021 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7081 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7141 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7201 gacgctcaag tcagag
//
