LOCUS       sequence-96311-e        8030 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yellow 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 8030)
  AUTHORS   Mazo-Vargas A, Park H, Aydin M, Buchler NE
  TITLE     ""Measuring fast gene dynamics in single cells with timelapse 
            luminescence microscopy""
  JOURNAL   Mol Biol Cell. 2014 Sep 17. pii: mbc.E14-07-1187.
  PUBMED    25232010
REFERENCE   2  (bases 1 to 8030)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pNB756
FEATURES             Location/Qualifiers
     source          1..8030
                     /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"
     primer_bind     complement(3809..3827)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      5478..5665
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5533..5557)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6035..6054)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6036..6054)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6064..6081)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6064..6080)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6073..6095)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6225..6680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6312..6331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6522..6543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6811..7614)
                     /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(7615..7835)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7861..7880)
                     /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 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 gcatggaaga cgccaaaaac ataaagaaag gcccggcgcc
     3781 attctatcct ctagaggatg gaaccgctgg agagcaactg cataaggcta tgaagagata
     3841 cgccctggtt cctggaacaa ttgcttttac agatgcacat atcgaggtga acatcacgta
     3901 cgcggaatac ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa
     3961 tacaaatcac agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt
     4021 gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga
     4081 attgctcaac agtatgaaca tttcgcagcc taccgtagtg tttgtttcca aaaaggggtt
     4141 gcaaaaaatt ttgaacgtgc aaaaaaaatt accaataatc cagaaaatta ttatcatgga
     4201 ttctaaaacg gattaccagg gatttcagtc gatgtatacg ttcgtcacat ctcatctacc
     4261 tcccggtttt aatgaatacg attttgtacc agagtccttt gatcgtgaca aaacaattgc
     4321 actgataatg aattcctctg gatctactgg gttacctaag ggtgtggccc ttccgcatag
     4381 aactgcctgc gtcagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc
     4441 ggatactgcg attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact
     4501 cggatatttg atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt
     4561 tttacgatcc cttcaggatt acaaaattca aagtgcgttg ctagtaccaa ccctattttc
     4621 attcttcgcc aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc
     4681 ttctgggggc gcacctcttt cgaaagaagt cggggaagcg gttgcaaaac gcttccatct
     4741 tccagggata cgacaaggat atgggctcac tgagactaca tcagctattc tgattacacc
     4801 cgagggggat gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt
     4861 tgtggatctg gataccggga aaacgctggg cgttaatcag agaggcgaat tatgtgtcag
     4921 aggacctatg attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga
     4981 caaggatgga tggctacatt ctggagacct tgcttactgg gacgaagacg aacacttctt
     5041 catagttggg cgcttgaagt ctttaatcaa atacaaaggt tatcaggtgg cccccgctga
     5101 attggaatcg atattgttac aacaccccaa catcttcgac gcgggcgtgg caggtcttcc
     5161 cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat
     5221 gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg
     5281 cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt accggaaaaa gagacgcaag
     5341 aaaaatcaga gagatcctca taaaggccaa gaagggaggt ggcagtatgt acaaaactag
     5401 tgcagcgaac gacgaaaatt acgcccttgc agcgtaataa ctagttaata caaataaggc
     5461 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     5521 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     5581 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     5641 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
     5701 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
     5761 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
     5821 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
     5881 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
     5941 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     6001 cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     6061 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     6121 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     6181 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc cctgtagcgg
     6241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6301 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6361 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6421 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6541 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6601 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6661 aatattaacg tttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     6721 tcacaccgca tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
     6781 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
     6841 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
     6901 aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt
     6961 tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat
     7021 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
     7081 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
     7141 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
     7201 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
     7261 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
     7321 accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc
     7381 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
     7441 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     7501 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     7561 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     7621 tcttcgtttc ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     7681 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     7741 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaagaaacc gaaatcaaaa
     7801 aaaagaataa aaaaaaaatg atgaattgaa ttgaaaagct gtggtatggt gcactctcag
     7861 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     7921 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     7981 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
