LOCUS       sequence-96362-e        6802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase (Promega) driven by SIC1 promoter.  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6802)
  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 6802)
  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: pNB767
FEATURES             Location/Qualifiers
     source          1..6802
                     /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"
     CDS             2568..4217
                     /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(2634..2652)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      4250..4437
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4305..4329)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4807..4826)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4808..4826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4836..4853)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4836..4852)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4845..4867)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4997..5452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5084..5103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5294..5315
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5583..6386)
                     /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(6387..6607)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6633..6652)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6752..6774
                     /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 ctggagctcc
     2281 tccggcgatc aaacatacca ttcccccctc aaatcaatat gattaagtca cgataatatc
     2341 tgcactattg ccgtgtttcc ttctccatcg acagcgtcga tcctgtctat tatagccagc
     2401 acactgaaaa tttcaatggc tggcttgttg ttcgcgtcgc aacttttgat atttataaaa
     2461 agagagaagg catatccaaa ggataaggta ttgtctttgt aaatcaatca accaaacctc
     2521 tacggaattt tgacccttga agcagggact attacacgtt aattaacatg gaagacgtca
     2581 aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc gctggagagc
     2641 aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg
     2701 cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt cggttggcag
     2761 aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact
     2821 ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg
     2881 cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg cagcctaccg
     2941 tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa
     3001 tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt
     3061 acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt gtgccagagt
     3121 ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct actggtctgc
     3181 ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat gccagagatc
     3241 ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc
     3301 acggttttgg aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa
     3361 tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag attcaaagtg
     3421 cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt gacaaatacg
     3481 atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag gaagtcgggg
     3541 aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg ctcactgaga
     3601 ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag
     3661 ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta
     3721 atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat gtaaacaatc
     3781 cggaagcgac caacgccttg attgacaagg atggatggct acattctgga gacatagctt
     3841 actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg attaagtaca
     3901 aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac cccaacatct
     3961 tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg
     4021 ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccggtc
     4081 aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag
     4141 gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg
     4201 gcggaaagat cgccgtgtct cgagaataag gcgcgccact tctaaataag cgaatttctt
     4261 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     4321 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     4381 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag
     4441 atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc ccgccgggtc
     4501 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
     4561 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
     4621 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
     4681 agacctgcga gcagggaaac gctcccctca cagacgcgtg tacgcatgta acattatact
     4741 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc
     4801 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     4861 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     4921 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4981 aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5041 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5101 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5161 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5221 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5281 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5341 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5401 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcctgatgc
     5461 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat
     5521 aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt
     5581 ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt
     5641 tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg
     5701 tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc
     5761 ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca
     5821 cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca
     5881 acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac
     5941 atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca
     6001 atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat
     6061 acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg
     6121 aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg
     6181 gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct
     6241 tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt
     6301 tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca
     6361 cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg
     6421 tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata
     6481 tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa
     6541 tcaaaaaaat ttcaaagaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg
     6601 aattgaaaag ctgtggtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6661 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6721 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6781 ccgtcatcac cgaaacgcgc ga
//
