LOCUS       sequence-448242-        5239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of luciferase driven by mPer2 promoter fragment 
            containing mutated E-box2 (bases -112 to +98 with respect to 
            transcription start site at +1) and SV40 promoter (backbone).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5239)
  AUTHORS   Yoo SH, Ko CH, Lowrey PL, Buhr ED, Song EJ, Chang S, Yoo OJ, 
            Yamazaki S, Lee C, Takahashi JS
  TITLE     A noncanonical E-box enhancer drives mouse Period2 circadian 
            oscillations in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Feb 15;102(7):2608-13. Epub 2005 Feb 
            7.
  PUBMED    15699353
REFERENCE   2  (bases 1 to 5239)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2005-02-15"; volume: "102(7)"; pages: 
            "2608-13. Epub 2005 Feb 7"
COMMENT     Sequence Label: pGL3 Pro Mut E2
FEATURES             Location/Qualifiers
     source          1..5239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    587..635
                     /note="synthetic polyadenylation signal"
     misc_feature    649..740
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     689..708
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1024..1220
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1071..1206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1250..1259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1256..2908
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1322..1340)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2949..3070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2960..2979)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3014..3033
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3318..3335)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3489..4077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3569..3588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4248..5039,5040..5108))
                     /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     4871..4890
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5109..5213)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
      541 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
      601 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
      661 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
      721 aggtgccaga acatttctct atcgataggt accgagctct tacgcgtgct agccccgcgg
      781 atccgagcgt agctctcagg ttccgccccg ccagtatgca aatgaggtgg cactccgacc
      841 aatggcgcgc gcaggggcgg gctcagcgcg cgcggtgcta gtttccacta tgtgacagcg
      901 gagggcgacg cggcggcagc ggcgctactg ggactagcgg ctccgggcgg ctgcggcgca
      961 ggccgagcgc accaagtgac gggccgagca agggatctag agcagggctc gagatctgcg
     1021 atctgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     1081 taactccgcc cagttccgcc cattctccgc cccatcgctg actaattttt tttatttatg
     1141 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     1201 gaggcctagg cttttgcaaa aagcttggca ttccggtact gttggtaaag ccaccatgga
     1261 agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc
     1321 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
     1381 tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg
     1441 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
     1501 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
     1561 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca
     1621 gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
     1681 gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca
     1741 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
     1801 gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac
     1861 tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc
     1921 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
     1981 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
     2041 cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat
     2101 tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga
     2161 caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga
     2221 agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct
     2281 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     2341 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     2401 gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt
     2461 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     2521 catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat
     2581 taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc
     2641 caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc
     2701 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     2761 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     2821 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     2881 caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc
     2941 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3001 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3061 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3121 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc
     3181 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     3241 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     3301 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3361 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3421 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3661 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3901 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4141 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4201 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4261 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4321 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4381 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4441 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4501 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4561 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4621 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4681 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4741 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4801 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4861 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4921 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4981 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5041 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5101 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5221 ccccgaaaag tgccacctg
//
