LOCUS       sequence-96349-e        8115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green beetle luciferase with nuclear localization signal (NLS) and 
            adjustable degron tag (ssrA), driven by LYS9 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8115)
  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 8115)
  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: pNB754
FEATURES             Location/Qualifiers
     source          1..8115
                     /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"
     primer_bind     3758..3774
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3758..3774
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3801..3821
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5563..5750
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5618..5642)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6120..6139)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6121..6139)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6149..6166)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6149..6165)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6158..6180)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6310..6765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6397..6416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6607..6628
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6896..7699)
                     /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(7700..7920)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7946..7965)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8065..8087
                     /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 ctggagctca
     2281 agaagcccac gagcaataat cctcctggca tatttactaa tggaacacgt tctcatagaa
     2341 agcaacccct aaaaagaaag gactagtcaa ccacaagcct tttaaaaaga aaaattgaaa
     2401 aaataataaa aaaatttgaa aaagaaaaaa aaaaatcata aaaccaatct taagaaaaac
     2461 caaaaaaaaa ccgctaatta aaatctaaaa aattgaaaca agaaactgta tgattagtaa
     2521 ctaatatact ggcgcggaag ggaatgaatg ccatttaaat gagatgatta tgaaaatata
     2581 gtgtagagta gatgcgcgaa accgaggcag aaaagaaaaa gagacaagtt aaaaattgaa
     2641 tataatgata tatgggcact ttcttctgtg ctaggaaacg gaggtagaag gggatagcaa
     2701 gcttgagcag taattaaaaa aaaaaaaaaa gcatattgat agatagctca tttgtagtta
     2761 actgacgaca tttatgtaca taattaaaat agaagaaaat tttcaaaaaa aattctcctg
     2821 gtgattgtta aggacaagaa aacaagccat gttgtaacca ataaatgtta cacgccttca
     2881 aaataaaaaa aagatggaca aaagagataa atcttttgct ttttcttact ctgtgttgtt
     2941 gtcaacaaag acaacaaacg tgtgaataat ctacgggatt cgtttaccat aggctaccta
     3001 gaaatggcat agtcatggag ataaagtccc atgatattgt aaactaaatt gaaagtaaaa
     3061 tcaaggaaaa aacaagtgag attgcaataa gcaatcaaga gagaaaagtt ttttgactga
     3121 ttttcgactt gcttttatac cgattttgaa aagtcgcggg gaaaagctgg caattatttt
     3181 tcattttttt tcattagggc tagaagttgc taagagcaaa caaactgtta cttgaaaggt
     3241 tttcccacgt tggtagctac atatgcaaga gtcttatgta tcgtatctaa gtgccacgta
     3301 ggggattccc atcatttgat gatttccaaa tataatacct gtagagagcg gtggagcaaa
     3361 agtcaaattt taatcgcaac tgcagacaag tcaagctgag gaaattgtgg atgatctctt
     3421 gtttcttttg atattcacca caacagaagt gaagagtgtg attgcggtta ctactgacca
     3481 cgaagcaatg cgtttagtag tgaaaagaat tactcatact ctggaatcga aattccgttg
     3541 gaaaaattcg ctttgtagtg aaaaataaag atgtcaataa agggtattga gaatttccaa
     3601 tggaattatc agcaatagat gatagaaagt agcacagaat ttggcttaat ggtatataaa
     3661 ccgtagggtc ctggtaaaat tacatgggaa ggatccttag gcagtaggga aaacttatca
     3721 ggacaattga gttatattaa cgtattatat attttaatct agaactagtg gatcccccgg
     3781 gttaattaac catggctcca ccaaaaaaaa aaagaaaagt tgccatgggt gggggcagca
     3841 tgatgaagag agaaaaaaat gttgtttacg gccccgaacc caaacaccct ttgggaaact
     3901 ttactgctgg agaaatgctc tacaatgccc ttcataagca ttcccacata ccgcaagcaa
     3961 tattagatgt gatgggtaat gaatcgctgt catatcaaga atttttcgac actactgtca
     4021 agctaggaca aagtcttcaa aattgtggat acaagatgaa tgatgtagtg tcgatctgtg
     4081 ctgagaacaa taaaagattt ttcatcccca ttatttcagc ttggtatatt ggtatggttg
     4141 tagcacctgt taatgaagac tacatcccag atgaactttg taaagtcacg ggtatatcaa
     4201 aaccaatact ggtcttcact acaagaaaaa tcttacctaa ggtattagag gtaaaggaca
     4261 gaactaatta cataaagaga attataatac tagattctga agaaaatctg cttggttgcg
     4321 aaagtcttca taattttatg tcccgttatt cggataataa tcttcagacc tttaagcctc
     4381 tacattacga tcctgttgat caagtggcag ctattttatg ttcgtcaggc acaactggat
     4441 tacccaaagg tgttatgcaa acgcatagaa acatttgtgt tcgacttaca catgcttcgg
     4501 atcccagagt tggaacacaa cttattcctg gagtatcagt tttggcgtat ctgcctttct
     4561 tccatgcttt tgggttttct ataaacttag gatactttat ggtgggcctt cgtgttgtta
     4621 tgctaagacg atttaatcaa gaagtatttt taaaagccat tcaagattat gaagttcgaa
     4681 gtgtaatcaa cgttccatca acaatactgt tcttgtcgaa aagtccttta gttgacaaat
     4741 acgatttatc gactttggcg gaattgtgtt gcggcgctgc accattagca aaggaagttg
     4801 ctgagatagc agtgaaacga ctaaacctgc caggaattcg ttgtggatat ggtttgacag
     4861 agtctacttc agctaatata catactcttc acaatgaatt taagtcagga tcacttggaa
     4921 aagtcactcc ttatatggct gcgaaaataa tagataggaa cactggtgaa gctttgggac
     4981 caaatcaagt tggagaacta tgcatctggg gtcctatggt aacaaaaggt tacgtgaaca
     5041 atccacaagc caccaaagag gctattgatg acgacggttg gcttcactct ggagactttg
     5101 gatactatga tgaggatgaa tatttctata tagtggaccg ttacaaggaa cttattaaat
     5161 ataaaggcta tcaggtagca cctgtagaat tagaagagat tttattacaa catccaggta
     5221 taagagatgt tgctgtcgtt ggtattcctg atatagaagc tggagaacta ccagctgggt
     5281 tcgtggttaa acaacccgga gcacaactta cagcaaaaga agtttacgat tttcttgccc
     5341 aacgggtctc tcattcaaag tatttgcgtg gaggagttcg attcgttgat tcaataccca
     5401 ggaatgttac aggtaaaatt tcaagaaaag aacttcgaga ggcgttgatg gaaaaagctg
     5461 gaggtggcag tatgtacaaa actagtgcag cgaacgacga aaattacgcc cttgcagcgt
     5521 aataactagt taatacaaat aaggcgcgcc acttctaaat aagcgaattt cttatgattt
     5581 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     5641 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     5701 ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct
     5761 agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc
     5821 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     5881 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     5941 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     6001 cgagcaggga aacgctcccc tcacagacgc gtgtacgcat gtaacattat actgaaaacc
     6061 ttgcttgaga aggttttggg acgctcgaag gctttaattt gcggccggta cccaattcgc
     6121 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     6181 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     6241 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     6301 aatggcgcga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6361 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6421 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6481 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6541 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6601 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6661 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6721 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcctga tgcggtattt
     6781 tctccttacg catctgtgcg gtatttcaca ccgcataggg taataactga tataattaaa
     6841 ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt
     6901 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     6961 ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc
     7021 ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat
     7081 ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt
     7141 ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac
     7201 ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa
     7261 ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg
     7321 ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg
     7381 cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta
     7441 aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat
     7501 cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta
     7561 actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt
     7621 cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct
     7681 tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt
     7741 gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca
     7801 atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa
     7861 aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa
     7921 aagctgtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     7981 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     8041 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     8101 caccgaaacg cgcga
//
