LOCUS       sequence-96326-e        7501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase (Promega) driven by ADE17 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7501)
  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 7501)
  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: pNB761
FEATURES             Location/Qualifiers
     source          1..7501
                     /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     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             3267..4916
                     /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(3333..3351)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      4949..5136
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5004..5028)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5506..5525)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5507..5525)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5535..5552)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5535..5551)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5544..5566)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5696..6151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5783..5802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5993..6014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6282..7085)
                     /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(7086..7306)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7332..7351)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7451..7473
                     /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 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 attaacatgg aagacgtcaa aaacataaag aaaggcccgg
     3301 cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga
     3361 gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca
     3421 cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc
     3481 tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg
     3541 tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac
     3601 gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg
     3661 ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca
     3721 tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc
     3781 tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa
     3841 ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc
     3901 atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca
     3961 ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta
     4021 cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc
     4081 tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat
     4141 tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa
     4201 ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc
     4261 atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta
     4321 cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga
     4381 aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg
     4441 tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga
     4501 ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact
     4561 tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag gtggctcccg
     4621 ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc
     4681 ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga
     4741 cgatgacgga aaaagagatc gtggattacg tcgccggtca agtaacaacc gcgaaaaagt
     4801 tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg
     4861 caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc gccgtgtctc
     4921 gagaataagg cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat
     4981 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     5041 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     5101 gtatgaggtc gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt
     5161 aatatagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg
     5221 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
     5281 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
     5341 gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg
     5401 ctcccctcac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     5461 tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg
     5521 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5581 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5641 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcgacgcg
     5701 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5761 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5821 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5881 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5941 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6001 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6061 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6121 aattttaaca aaatattaac gtttacaatt tcctgatgcg gtattttctc cttacgcatc
     6181 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt
     6241 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     6301 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     6361 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     6421 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     6481 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     6541 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     6601 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     6661 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     6721 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     6781 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     6841 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     6901 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     6961 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     7021 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     7081 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     7141 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     7201 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaagaaac
     7261 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga attgaaaagc tgtggtatgg
     7321 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     7381 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     7441 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     7501 a
//
