LOCUS       sequence-444692-        5047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of luciferase driven by mPer2 promoter fragment 
            containing E-box2 (bases -112 to +98 with respect to transcription 
            start site at +1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5047)
  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 5047)
  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 basic E2
FEATURES             Location/Qualifiers
     source          1..5047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    143..191
                     /note="synthetic polyadenylation signal"
     misc_feature    205..296
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     245..264
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..2272
                     /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(686..704)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2313..2434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2324..2343)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2378..2397
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2682..2699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2853..3441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2933..2952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3612..4403,4404..4472))
                     /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     4235..4254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4473..4577)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4604..5032
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4691..4710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4901..4922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgcttacaat ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
       61 gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac
      121 gggaggtact tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
      181 ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa
      241 caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg
      301 ataggtaccg agctcttacg cgtgctagcc ccgcggatcc gagcgtagct ctcaggttcc
      361 gccccgccag tatgcaaatg aggtggcact ccgaccaatg gcgcgcgcag gggcgggctc
      421 agcgcgcgcg gtcacgtttt ccactatgtg acagcggagg gcgacgcggc ggcagcggcg
      481 ctactgggac tagcggctcc gggcggctgc ggcgcaggcc gagcgcacca agtgacgggc
      541 cgagcaaggg atctagagca gggctcgaga tctgcgatct aagtaagctt ggcattccgg
      601 tactgttggt aaagccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt
      661 ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc
      721 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc
      781 tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac
      841 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg
      901 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt
      961 gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca
     1021 aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc
     1081 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc
     1141 cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact
     1201 gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac
     1261 tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga
     1321 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg
     1381 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct
     1441 gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt
     1501 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc
     1561 tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc
     1621 aggtatcagg caaggatatg ggctcactga gactacatca gctattctga ttacacccga
     1681 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt
     1741 ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg
     1801 tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa
     1861 ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat
     1921 cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt
     1981 ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga
     2041 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac
     2101 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg
     2161 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa
     2221 aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagag
     2281 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2341 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2401 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2461 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2521 tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     2581 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     2641 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     2701 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2761 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2821 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2881 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2941 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3001 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3061 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3121 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3181 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3241 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3301 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3361 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3421 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3481 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3541 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3601 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3661 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3721 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3781 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3841 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3901 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3961 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4021 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4081 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4141 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4201 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4261 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4321 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4381 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4441 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4501 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4561 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     4621 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4681 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4741 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4801 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4861 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4921 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4981 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5041 atattaa
//
