LOCUS       sequence-96201-e        7031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yellow 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 7031)
  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 7031)
  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: pNB690
FEATURES             Location/Qualifiers
     source          1..7031
                     /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"
     primer_bind     complement(2810..2828)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      4479..4666
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4534..4558)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5036..5055)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5037..5055)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5065..5082)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5065..5081)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5074..5096)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5226..5681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5313..5332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5523..5544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5812..6615)
                     /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(6616..6836)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6862..6881)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 acgccaaaaa
     2761 cataaagaaa ggcccggcgc cattctatcc tctagaggat ggaaccgctg gagagcaact
     2821 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
     2881 tatcgaggtg aacatcacgt acgcggaata cttcgaaatg tccgttcggt tggcagaagc
     2941 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     3001 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     3061 cgacatttat aatgaacgtg aattgctcaa cagtatgaac atttcgcagc ctaccgtagt
     3121 gtttgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaaat taccaataat
     3181 ccagaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtatac
     3241 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtac cagagtcctt
     3301 tgatcgtgac aaaacaattg cactgataat gaattcctct ggatctactg ggttacctaa
     3361 gggtgtggcc cttccgcata gaactgcctg cgtcagattc tcgcatgcca gagatcctat
     3421 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     3481 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     3541 tagatttgaa gaagagctgt ttttacgatc ccttcaggat tacaaaattc aaagtgcgtt
     3601 gctagtacca accctatttt cattcttcgc caaaagcact ctgattgaca aatacgattt
     3661 atctaattta cacgaaattg cttctggggg cgcacctctt tcgaaagaag tcggggaagc
     3721 ggttgcaaaa cgcttccatc ttccagggat acgacaagga tatgggctca ctgagactac
     3781 atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt
     3841 tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca
     3901 gagaggcgaa ttatgtgtca gaggacctat gattatgtcc ggttatgtaa acaatccgga
     3961 agcgaccaac gccttgattg acaaggatgg atggctacat tctggagacc ttgcttactg
     4021 ggacgaagac gaacacttct tcatagttgg gcgcttgaag tctttaatca aatacaaagg
     4081 ttatcaggtg gcccccgctg aattggaatc gatattgtta caacacccca acatcttcga
     4141 cgcgggcgtg gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt
     4201 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     4261 aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     4321 taccggaaaa agagacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggagg
     4381 tggcagtatg tacaaaacta gtgcagcgaa cgacgaaaat tacgcccttg cagcgtaata
     4441 actagttaat acaaataagg cgcgccactt ctaaataagc gaatttctta tgatttatga
     4501 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     4561 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     4621 ctcaggtata gtatgaggtc gctcttattg accacacctc taccggcaga tccgctaggg
     4681 ataacagggt aatatagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc
     4741 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
     4801 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
     4861 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
     4921 cagggaaacg ctcccctcac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     4981 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta
     5041 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5101 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5161 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5221 gcgcgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5281 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5341 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5401 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5461 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5521 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5581 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5641 atttaacgcg aattttaaca aaatattaac gtttacaatt tcctgatgcg gtattttctc
     5701 cttacgcatc tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga
     5761 agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg
     5821 ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac
     5881 cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt
     5941 agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa
     6001 acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct
     6061 ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt
     6121 agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc
     6181 tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc
     6241 caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga
     6301 gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa
     6361 attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt
     6421 caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc
     6481 cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg
     6541 catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata
     6601 tgtagctttc gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt
     6661 taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct
     6721 aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt
     6781 tcaaagaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga attgaaaagc
     6841 tgtggtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     6901 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6961 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     7021 gaaacgcgcg a
//
