LOCUS       sequence-96202-e        7028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red beetle luciferase with nuclear localization signal (NLS) and 
            ClpXP degron tag (ssrA), driven by MET17 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7028)
  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 7028)
  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: pNB691
FEATURES             Location/Qualifiers
     source          1..7028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2241..2261
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2312..2661
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     2662..2678
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2662..2678
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2705..2725
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4476..4663
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4531..4555)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5033..5052)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5034..5052)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5062..5079)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5062..5078)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5071..5093)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5223..5678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5310..5329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5520..5541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5809..6612)
                     /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"
     promoter        complement(6613..6833)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6859..6878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6978..7000
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc
     2281 ggatgcaagg gttcgaatcc cttagctctc attatttttt gctttttctc ttgaggtcac
     2341 atgatcgcaa aatggcaaat ggcacgtgaa gctgtcgata ttggggaact gtggtggttg
     2401 gcaaatgact aattaagtta gtcaaggcgc catcctcatg aaaactgtgt aacataataa
     2461 ccgaagtgtc gaaaaggtgg caccttgtcc aattgaacac gctcgatgaa aaaaataaga
     2521 tatatataag gttaagtaaa gcgtctgtta gaaaggaagt ttttcctttt tcttgctctc
     2581 ttgtcttttc atctactatt tccttcgtgt aatacagggt cgtcagatac atagatacaa
     2641 ttctattacc cccatccata ctctagaact agtggatccc ccgggttaat taaccatggc
     2701 tccaccaaaa aaaaaaagaa aagttgccat gggtgggggc agcatggaag aagaaaacat
     2761 tgtgaatgga gatcgtcctc gtgatctagt ttttcctggc acagcaggac tacaattata
     2821 tcaatcatta tataaatatt catatattac tgacggaata atcgatgccc ataccaatga
     2881 agtaatatca tatgctcaaa tatttgaaac cagctgccgc ttggcagtta gtctagaaaa
     2941 atatggcttg gatcataaca atgttgtggc aatatgcagt gaaaacaaca tacacttttt
     3001 tggcccttta attgctgctt tataccaagg aataccaatg gcaacatcaa atgatatgta
     3061 tacagaaagg gagatgattg gccatttgaa tatatcgaaa ccatgcctta tgttttgttc
     3121 aaagaaatca ctcccattta ttctgaaagt acaaaaacat ctagatttcc ttaaaaaagt
     3181 catagtcatt gatagtatgt acgacatcaa tggcgttgaa tgcgtattta gctttgtttc
     3241 acgttatact gatcacgcct ttgatccagt gaaatttaac ccaaaagagt ttgatccctt
     3301 ggaaagaacc gcattaatta tgacatcatc tggaacaact ggattgccta aaggggtagt
     3361 aataagccat agaagtataa ctataagatt cgtccatagc agtgatccca tctatggtac
     3421 tcgtattgct ccagatacat caattcttgc tatagcaccg ttccatcatg cctttggact
     3481 gtttactgca ctagcttact ttccagtagg acttaagatt gtaatggtga agaaatttga
     3541 gggcgaattc ttcttaaaaa ccatacaaaa ttacaaaatc gcttctattg tagttcctcc
     3601 tccaattatg gtatatttgg ctaaaagtcc attagtcgat gaatacaatt tatcgagctt
     3661 aacggaaatt gcttgtggag ggtctccttt aggaagagac atcgcagata aagtagcaaa
     3721 gagattgaaa gtacatggaa tcctacaagg atatggatta accgaaacct gcagcgctct
     3781 aatacttagc cccaatgatc gagaacttaa aaaaggtgca attggaacgc ctatgccata
     3841 tgttcaagtt aaagttatag acatcaatac tgggaaggcg ctaggaccaa gagaaaaagg
     3901 cgaaatatgc ttcaaaagtc aaatgcttat gaaaggttat cacaacaatc cgcaagcaac
     3961 tcgtgatgct cttgacaaag atggttggct tcatactggg gatcttggat attacgacga
     4021 agacagattt atctatgtag ttgatcgatt gaaagaactt attaaatata aaggttatca
     4081 ggttgcgcct gctgaactgg aaaatctgct tttacaacat ccaaatattt ctgatgcggg
     4141 tgttattgga attccggacg aatttgctgg tcaattacct tccgcgtgtg ttgtgttaga
     4201 gcctggtaag acaatgaccg aaaaggaagt tcaggattat attgcagagc tagtcactac
     4261 aactaaacat cttcgaggcg gtgtcgtatt tatagatagt attccaaaag gcccaacagg
     4321 aaaactcatg agaaacgaac tccgtgcaat atttgcccgg gaacaggcaa aaggaggtgg
     4381 cagtatgtac aaaactagtg cagcgaacga cgaaaattac gcccttgcag cgtaataact
     4441 agttaataca aataaggcgc gccacttcta aataagcgaa tttcttatga tttatgattt
     4501 ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg
     4561 ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc
     4621 aggtatagta tgaggtcgct cttattgacc acacctctac cggcagatcc gctagggata
     4681 acagggtaat atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac
     4741 atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt
     4801 gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat
     4861 tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag
     4921 ggaaacgctc ccctcacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     4981 agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag
     5041 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     5101 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     5161 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     5221 cgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5281 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5341 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5401 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5461 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5521 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5581 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5641 taacgcgaat tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt
     5701 acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc
     5761 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     5821 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     5881 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     5941 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     6001 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     6061 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     6121 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     6181 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     6241 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     6301 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     6361 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     6421 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     6481 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     6541 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     6601 agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa
     6661 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     6721 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     6781 aagaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt gaaaagctgt
     6841 ggtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     6901 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     6961 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     7021 acgcgcga
//
