LOCUS       sequence-96305-e        8018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green beetle luciferase with nuclear localization signal (NLS) and 
            ClpXP degron tag (ssrA), driven by LEU1 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8018)
  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 8018)
  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: pNB753
FEATURES             Location/Qualifiers
     source          1..8018
                     /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"
     CDS             3184..3195
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3704..3724
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5466..5653
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5521..5545)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6023..6042)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6024..6042)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6052..6069)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6052..6068)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6061..6083)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6213..6668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6300..6319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6510..6531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6799..7602)
                     /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(7603..7823)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7849..7868)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7968..7990
                     /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 ctggagctct
     2281 gatgaacggt aagccaatga aggaaaagaa gtctaccaga agtgtcgaag acttcatggc
     2341 tgctatgcaa agagtctcta ctcaacacga aaaggaaacc taatcctgtt gaagtagcat
     2401 ttaatcataa tttttgtcac attttaatca acttgatttt tctggtttaa tttttctaat
     2461 tttaatttta atttttttat caatgggaac tgatacacta aaaagaatta ggagccaaca
     2521 agaataagcc gcttatttcc tactagagtt tgcttaaaat ttcatctcga attgtcattc
     2581 taatatttta tccacacaca caccttaaaa tttttagatt aaatggcatc aactcttagc
     2641 ttcacacaca cacacacacc gaagctggtt gttttatttg atttgatata attggtttct
     2701 ctggatggta ctttttcttt cttggttatt tcctatttta aaatatgaaa cgcacacaag
     2761 tcataattat tctaatagag cacaattcac aacacgcaca tttcaacttt aatatttttt
     2821 tagaaacact ttatttagtc taattcttaa tttttaatat atataatgca cacacactaa
     2881 tttattcatt aattttttat tgagtaggat ttgaaaatat ttggtatctt tgcaagatgt
     2941 ttgtatagag ggacaaagaa tcgtctttat tatggtcaag gctttacgtc ataatagttc
     3001 ctgcccagct cttctataat actttaaaga tctcttctcg tttgctccat ttggaagtct
     3061 cgcttacgtt tatgcgccca tacagacact caagatacac acttacatga acgtatacaa
     3121 atttactaac actacttgaa aatatgaacc acagtacatc atattaagac gtagtattcg
     3181 atgattgaag gccgcctccg cgaaatacct ttactgattt tgccggttaa tcgcatcgaa
     3241 atttcttcat cacaagaaag caaacaaatc gccaggccat tctacaagtt tccttttctt
     3301 atgaagatgt aaaagctact aaggcgtcat tactctagat gactcagttt agtctgacct
     3361 tctatagtat actaccctgg cgctatgatg atgagcggtt cttttattgc ggaaacgaaa
     3421 attccgggac cggcgaaatt tgcccggttt tgtccgtaac cggcttcatg agtcggcttc
     3481 aatagtagtt gaatacttat ttaaacagca gaacttaact cactcatcac gctgtttccg
     3541 ctgaattttc tcaaaatatc taagcagtca acaaatataa agaatattga aattgacagt
     3601 ttttgtcgct atcgattttt attatttgct gttttaaatc ttggtttaca ctccatccaa
     3661 actagaacta gtggatcccc cgggttaatt aaccatggct ccaccaaaaa aaaaaagaaa
     3721 agttgccatg ggtgggggca gcatgatgaa gagagaaaaa aatgttgttt acggccccga
     3781 acccaaacac cctttgggaa actttactgc tggagaaatg ctctacaatg cccttcataa
     3841 gcattcccac ataccgcaag caatattaga tgtgatgggt aatgaatcgc tgtcatatca
     3901 agaatttttc gacactactg tcaagctagg acaaagtctt caaaattgtg gatacaagat
     3961 gaatgatgta gtgtcgatct gtgctgagaa caataaaaga tttttcatcc ccattatttc
     4021 agcttggtat attggtatgg ttgtagcacc tgttaatgaa gactacatcc cagatgaact
     4081 ttgtaaagtc acgggtatat caaaaccaat actggtcttc actacaagaa aaatcttacc
     4141 taaggtatta gaggtaaagg acagaactaa ttacataaag agaattataa tactagattc
     4201 tgaagaaaat ctgcttggtt gcgaaagtct tcataatttt atgtcccgtt attcggataa
     4261 taatcttcag acctttaagc ctctacatta cgatcctgtt gatcaagtgg cagctatttt
     4321 atgttcgtca ggcacaactg gattacccaa aggtgttatg caaacgcata gaaacatttg
     4381 tgttcgactt acacatgctt cggatcccag agttggaaca caacttattc ctggagtatc
     4441 agttttggcg tatctgcctt tcttccatgc ttttgggttt tctataaact taggatactt
     4501 tatggtgggc cttcgtgttg ttatgctaag acgatttaat caagaagtat ttttaaaagc
     4561 cattcaagat tatgaagttc gaagtgtaat caacgttcca tcaacaatac tgttcttgtc
     4621 gaaaagtcct ttagttgaca aatacgattt atcgactttg gcggaattgt gttgcggcgc
     4681 tgcaccatta gcaaaggaag ttgctgagat agcagtgaaa cgactaaacc tgccaggaat
     4741 tcgttgtgga tatggtttga cagagtctac ttcagctaat atacatactc ttcacaatga
     4801 atttaagtca ggatcacttg gaaaagtcac tccttatatg gctgcgaaaa taatagatag
     4861 gaacactggt gaagctttgg gaccaaatca agttggagaa ctatgcatct ggggtcctat
     4921 ggtaacaaaa ggttacgtga acaatccaca agccaccaaa gaggctattg atgacgacgg
     4981 ttggcttcac tctggagact ttggatacta tgatgaggat gaatatttct atatagtgga
     5041 ccgttacaag gaacttatta aatataaagg ctatcaggta gcacctgtag aattagaaga
     5101 gattttatta caacatccag gtataagaga tgttgctgtc gttggtattc ctgatataga
     5161 agctggagaa ctaccagctg ggttcgtggt taaacaaccc ggagcacaac ttacagcaaa
     5221 agaagtttac gattttcttg cccaacgggt ctctcattca aagtatttgc gtggaggagt
     5281 tcgattcgtt gattcaatac ccaggaatgt tacaggtaaa atttcaagaa aagaacttcg
     5341 agaggcgttg atggaaaaag ctggaggtgg cagtatgtac aaaactagtg cagcgaacga
     5401 cgaaaattac gcccttgcag cgtaataact agttaataca aataaggcgc gccacttcta
     5461 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
     5521 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
     5581 actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc
     5641 acacctctac cggcagatcc gctagggata acagggtaat atagatctgt ttagcttgcc
     5701 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
     5761 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
     5821 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
     5881 tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgtgtacg
     5941 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
     6001 tttgcggccg gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
     6061 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     6121 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     6181 acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc tgtagcggcg cattaagcgc
     6241 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     6301 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     6361 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     6421 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     6481 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     6541 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     6601 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     6661 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     6721 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta tacatgcatt
     6781 tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg cttcccagcc
     6841 tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa atagtcctct
     6901 tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc tatactgttg
     6961 acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca accaatcgta
     7021 accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa aatctttgtc
     7081 gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg agcccttgca
     7141 tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt ctgccgcctg
     7201 cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca
     7261 ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac caatgtcagc
     7321 aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct ttagcggctt
     7381 aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta gtaaacaaat
     7441 tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa catccaatga
     7501 agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag caacaggact
     7561 aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc ttcgtttcct
     7621 gcaggttttt gttctgtgca gttgggttaa gaatactggg caatttcatg tttcttcaac
     7681 actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt cttccttctg
     7741 ttcggagatt accgaatcaa aaaaatttca aagaaaccga aatcaaaaaa aagaataaaa
     7801 aaaaaatgat gaattgaatt gaaaagctgt ggtatggtgc actctcagta caatctgctc
     7861 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     7921 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     7981 gtgtcagagg ttttcaccgt catcaccgaa acgcgcga
//
