LOCUS       sequence-96358-e        8025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase (Promega) driven by LYS9 promoter.  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8025)
  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 8025)
  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: pNB763
FEATURES             Location/Qualifiers
     source          1..8025
                     /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"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     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             3791..5440
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDVKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVAGQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3857..3875)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      5473..5660
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5528..5552)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6030..6049)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6031..6049)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6059..6076)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6059..6075)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6068..6090)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6220..6675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6307..6326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6517..6538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6806..7609)
                     /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(7610..7830)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7856..7875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7975..7997
                     /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 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat 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 atggaagacg tcaaaaacat aaagaaaggc ccggcgccat tctatccgct
     3841 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     3901 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt
     3961 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     4021 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     4081 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     4141 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt
     4201 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga
     4261 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     4321 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa
     4381 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt
     4441 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     4501 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     4561 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct
     4621 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa
     4681 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc
     4741 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag
     4801 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     4861 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     4921 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat
     4981 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     5041 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg
     5101 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat
     5161 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc
     5221 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     5281 gatcgtggat tacgtcgccg gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     5341 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     5401 gatcctcata aaggccaaga agggcggaaa gatcgccgtg tctcgagaat aaggcgcgcc
     5461 acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     5521 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     5581 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt
     5641 attgaccaca cctctaccgg cagatccgct agggataaca gggtaatata gatctgttta
     5701 gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct
     5761 tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc
     5821 catacatccc catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag
     5881 caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc
     5941 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     6001 gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     6061 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     6121 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6181 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat
     6241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     6661 taacgtttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     6721 ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     6781 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     6841 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     6901 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     6961 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     7021 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     7081 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     7141 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     7201 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     7261 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     7321 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     7381 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     7441 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     7501 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     7561 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     7621 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     7681 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     7741 ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag
     7801 aataaaaaaa aaatgatgaa ttgaattgaa aagctgtggt atggtgcact ctcagtacaa
     7861 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     7921 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     7981 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
