LOCUS       sequence-96189-e        7019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green 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 7019)
  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 7019)
  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: pNB689
FEATURES             Location/Qualifiers
     source          1..7019
                     /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      4467..4654
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4522..4546)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5024..5043)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5025..5043)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5053..5070)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5053..5069)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5062..5084)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5214..5669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5301..5320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5511..5532
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5800..6603)
                     /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(6604..6824)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6850..6869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6969..6991
                     /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 agcatgatga agagagaaaa
     2761 aaatgttgtt tacggccccg aacccaaaca ccctttggga aactttactg ctggagaaat
     2821 gctctacaat gcccttcata agcattccca cataccgcaa gcaatattag atgtgatggg
     2881 taatgaatcg ctgtcatatc aagaattttt cgacactact gtcaagctag gacaaagtct
     2941 tcaaaattgt ggatacaaga tgaatgatgt agtgtcgatc tgtgctgaga acaataaaag
     3001 atttttcatc cccattattt cagcttggta tattggtatg gttgtagcac ctgttaatga
     3061 agactacatc ccagatgaac tttgtaaagt cacgggtata tcaaaaccaa tactggtctt
     3121 cactacaaga aaaatcttac ctaaggtatt agaggtaaag gacagaacta attacataaa
     3181 gagaattata atactagatt ctgaagaaaa tctgcttggt tgcgaaagtc ttcataattt
     3241 tatgtcccgt tattcggata ataatcttca gacctttaag cctctacatt acgatcctgt
     3301 tgatcaagtg gcagctattt tatgttcgtc aggcacaact ggattaccca aaggtgttat
     3361 gcaaacgcat agaaacattt gtgttcgact tacacatgct tcggatccca gagttggaac
     3421 acaacttatt cctggagtat cagttttggc gtatctgcct ttcttccatg cttttgggtt
     3481 ttctataaac ttaggatact ttatggtggg ccttcgtgtt gttatgctaa gacgatttaa
     3541 tcaagaagta tttttaaaag ccattcaaga ttatgaagtt cgaagtgtaa tcaacgttcc
     3601 atcaacaata ctgttcttgt cgaaaagtcc tttagttgac aaatacgatt tatcgacttt
     3661 ggcggaattg tgttgcggcg ctgcaccatt agcaaaggaa gttgctgaga tagcagtgaa
     3721 acgactaaac ctgccaggaa ttcgttgtgg atatggtttg acagagtcta cttcagctaa
     3781 tatacatact cttcacaatg aatttaagtc aggatcactt ggaaaagtca ctccttatat
     3841 ggctgcgaaa ataatagata ggaacactgg tgaagctttg ggaccaaatc aagttggaga
     3901 actatgcatc tggggtccta tggtaacaaa aggttacgtg aacaatccac aagccaccaa
     3961 agaggctatt gatgacgacg gttggcttca ctctggagac tttggatact atgatgagga
     4021 tgaatatttc tatatagtgg accgttacaa ggaacttatt aaatataaag gctatcaggt
     4081 agcacctgta gaattagaag agattttatt acaacatcca ggtataagag atgttgctgt
     4141 cgttggtatt cctgatatag aagctggaga actaccagct gggttcgtgg ttaaacaacc
     4201 cggagcacaa cttacagcaa aagaagttta cgattttctt gcccaacggg tctctcattc
     4261 aaagtatttg cgtggaggag ttcgattcgt tgattcaata cccaggaatg ttacaggtaa
     4321 aatttcaaga aaagaacttc gagaggcgtt gatggaaaaa gctggaggtg gcagtatgta
     4381 caaaactagt gcagcgaacg acgaaaatta cgcccttgca gcgtaataac tagttaatac
     4441 aaataaggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
     4501 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     4561 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
     4621 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
     4681 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
     4741 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
     4801 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
     4861 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
     4921 cccctcacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     4981 tgggacgctc gaaggcttta atttgcggcc ggtacccaat tcgccctata gtgagtcgta
     5041 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5101 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5161 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc
     5221 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5281 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5341 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5401 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5461 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5521 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5581 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5641 ttttaacaaa atattaacgt ttacaatttc ctgatgcggt attttctcct tacgcatctg
     5701 tgcggtattt cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg
     5761 tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct
     5821 tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct
     5881 tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc
     5941 atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg
     6001 tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa
     6061 gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc
     6121 agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt
     6181 tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg
     6241 tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt
     6301 gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc
     6361 ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac
     6421 atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt
     6481 ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa
     6541 tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga
     6601 catgatttat cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg
     6661 gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc
     6721 cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg
     6781 aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg
     6841 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     6901 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6961 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
