LOCUS       sequence-96323-e        7591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green beetle luciferase with nuclear localization signal (NLS) and 
            adjustable degron tag (ssrA), driven by ADE17 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7591)
  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 7591)
  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: pNB752
FEATURES             Location/Qualifiers
     source          1..7591
                     /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     3234..3250
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3234..3250
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3277..3297
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5039..5226
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5094..5118)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5596..5615)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5597..5615)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5625..5642)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5625..5641)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5634..5656)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5786..6241
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5873..5892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6083..6104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6372..7175)
                     /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(7176..7396)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7422..7441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7541..7563
                     /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 ctggagctcg
     2281 ccgctagacc aactagacca gacaaggcta ggagactagg ctacaaagcc aagcaaggtt
     2341 tcgttatcta ccgtgtcagg gttagacgcg gtaacagaaa gagacctgtt ccaaagggtg
     2401 ctacttacgg taagccaact aaccaaggtg tcaatgaatt gaaataccaa agatccttga
     2461 gagctaccgc tgaagaaaga gttggccgtc gtgctgctaa cttgagagtc ttgaactcct
     2521 actgggtcaa ccaagactct acttacaagt acttcgaagt tatattggtt gatcctcaac
     2581 acaaggctat cagaagagat gctcggtaca actggatctg caatcccgtc cataagcatc
     2641 gtgaagctag aggcttgact gccaccggta agaaatccag gggtatcaac aagggacaca
     2701 aattcaacaa caccaaggcc ggcagaagaa agacctggaa aagacaaaat actctgtcgt
     2761 tatggaggta tagaaaatga aatagttaga accgtatata gatcaataat aatgcactat
     2821 gaacattttt tcaacctgat tacttcatcg ttccttttct tcgacttatt ttttttttat
     2881 tattcttcgc tctttggcag catacgcaat ttgaatttgc agtttttgac tcttttttcc
     2941 ttagcggaac gaacaatatt aatcatgcca actaagtcat tgcagaagtc aacttatcat
     3001 ttataaagaa gatcctaccc tacaatgtgt gtaccctgat gtgcccagaa atgaattgac
     3061 tcctctcata acttgttaat attcggatgt gcatatcacg ttcggatcta tgtatataaa
     3121 tgtacgctcg ttttaacctt tagtgtagta cctaaaatct ttcttaccag caactaatag
     3181 cccttgaagt agttttgcta gcttggacat caaagcacat atcaccatca aattctagaa
     3241 ctagtggatc ccccgggtta attaaccatg gctccaccaa aaaaaaaaag aaaagttgcc
     3301 atgggtgggg gcagcatgat gaagagagaa aaaaatgttg tttacggccc cgaacccaaa
     3361 caccctttgg gaaactttac tgctggagaa atgctctaca atgcccttca taagcattcc
     3421 cacataccgc aagcaatatt agatgtgatg ggtaatgaat cgctgtcata tcaagaattt
     3481 ttcgacacta ctgtcaagct aggacaaagt cttcaaaatt gtggatacaa gatgaatgat
     3541 gtagtgtcga tctgtgctga gaacaataaa agatttttca tccccattat ttcagcttgg
     3601 tatattggta tggttgtagc acctgttaat gaagactaca tcccagatga actttgtaaa
     3661 gtcacgggta tatcaaaacc aatactggtc ttcactacaa gaaaaatctt acctaaggta
     3721 ttagaggtaa aggacagaac taattacata aagagaatta taatactaga ttctgaagaa
     3781 aatctgcttg gttgcgaaag tcttcataat tttatgtccc gttattcgga taataatctt
     3841 cagaccttta agcctctaca ttacgatcct gttgatcaag tggcagctat tttatgttcg
     3901 tcaggcacaa ctggattacc caaaggtgtt atgcaaacgc atagaaacat ttgtgttcga
     3961 cttacacatg cttcggatcc cagagttgga acacaactta ttcctggagt atcagttttg
     4021 gcgtatctgc ctttcttcca tgcttttggg ttttctataa acttaggata ctttatggtg
     4081 ggccttcgtg ttgttatgct aagacgattt aatcaagaag tatttttaaa agccattcaa
     4141 gattatgaag ttcgaagtgt aatcaacgtt ccatcaacaa tactgttctt gtcgaaaagt
     4201 cctttagttg acaaatacga tttatcgact ttggcggaat tgtgttgcgg cgctgcacca
     4261 ttagcaaagg aagttgctga gatagcagtg aaacgactaa acctgccagg aattcgttgt
     4321 ggatatggtt tgacagagtc tacttcagct aatatacata ctcttcacaa tgaatttaag
     4381 tcaggatcac ttggaaaagt cactccttat atggctgcga aaataataga taggaacact
     4441 ggtgaagctt tgggaccaaa tcaagttgga gaactatgca tctggggtcc tatggtaaca
     4501 aaaggttacg tgaacaatcc acaagccacc aaagaggcta ttgatgacga cggttggctt
     4561 cactctggag actttggata ctatgatgag gatgaatatt tctatatagt ggaccgttac
     4621 aaggaactta ttaaatataa aggctatcag gtagcacctg tagaattaga agagatttta
     4681 ttacaacatc caggtataag agatgttgct gtcgttggta ttcctgatat agaagctgga
     4741 gaactaccag ctgggttcgt ggttaaacaa cccggagcac aacttacagc aaaagaagtt
     4801 tacgattttc ttgcccaacg ggtctctcat tcaaagtatt tgcgtggagg agttcgattc
     4861 gttgattcaa tacccaggaa tgttacaggt aaaatttcaa gaaaagaact tcgagaggcg
     4921 ttgatggaaa aagctggagg tggcagtatg tacaaaacta gtgcagcgaa cgacgaaaat
     4981 tacgcccttg cagcgtaata actagttaat acaaataagg cgcgccactt ctaaataagc
     5041 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
     5101 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
     5161 cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg accacacctc
     5221 taccggcaga tccgctaggg ataacagggt aatatagatc tgtttagctt gcctcgtccc
     5281 cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg
     5341 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
     5401 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct
     5461 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgtgt acgcatgtaa
     5521 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg
     5581 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     5641 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5701 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5761 cgcagcctga atggcgaatg gcgcgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5821 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5881 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5941 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6001 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6061 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6121 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6181 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     6241 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat
     6301 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     6361 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     6421 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     6481 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     6541 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     6601 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     6661 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     6721 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     6781 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     6841 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     6901 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     6961 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     7021 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     7081 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     7141 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     7201 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     7261 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     7321 attaccgaat caaaaaaatt tcaaagaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     7381 gatgaattga attgaaaagc tgtggtatgg tgcactctca gtacaatctg ctctgatgcc
     7441 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7501 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     7561 aggttttcac cgtcatcacc gaaacgcgcg a
//
