LOCUS       sequence-210773-        7917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Per2 transcription luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7917)
  AUTHORS   Mei L, Fan Y, Lv X, Welsh DK, Zhan C, Zhang EE
  TITLE     Long-term in vivo recording of circadian rhythms in brains of freely
            moving mice.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Apr 2. pii: 1717735115. doi: 
            10.1073/pnas.1717735115.
  PUBMED    29610316
REFERENCE   2  (bases 1 to 7917)
  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. 2018 Apr 2. pii: 1717735115. doi: 
            10.1073/pnas.1717735115."
COMMENT     Sequence Label: AAV-P(Per2)-DIO-intron2-dLUC
FEATURES             Location/Qualifiers
     source          1..7917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(689..722)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(773..806)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(812..931)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     CDS             complement(935..982)
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             complement(989..2635)
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             complement(2645..2665)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2690..2734)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(2756..2821)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3548..3571)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3571..3580
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3618..3636)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3619..3636)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(3642..3660)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3703..3722)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3705..3837
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    3902..3935
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3986..4019
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4044..4632
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4097..4117)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4515..4526)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4664..5140
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4863..4882)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5180..5320
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5180..5309
                     /label=AAV2 ITR
     rep_origin      5395..5850
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5482..5501)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5692..5713
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5867..5886)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5986..6008
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6046..6064)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6132..6236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6237..6305,6306..7097)
                     /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(6455..6474)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7268..7856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7757..7776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcggatt accgaggctg gtcacgtcgt cgcaggtgat
      181 aggccggggg ccctggtctc tgccggctgt gagttgcgca gcggccaagc accattcccc
      241 cgcgccgcag tggtacgcgc cactccgggg ctgcacgagc gggccaccgc cgtgccaggt
      301 gaatggaagt cccgcaggcc ggaagtggac gagcctactc gcccgggcgc gggggggcgc
      361 aagagcgcgc agcatcttca ttgaggaacc cgggcggcga acatggagtt ccatgtgcgt
      421 cttatgtaaa gagagcgacg ggcgtctcca ccaattgacg agcgtagctc tcaggttccg
      481 ccccgccagt atgcaaatga ggtggcactc cgaccaatgg cgcgcgcagg ggcgggctca
      541 gcgcgcgcgg tcacgttttc cactatgtga cagcggaggg cgacgcggcg gcagcggcgc
      601 tactgggact agcggctccg ggcggctgcg gcgcaggccg agcgcaccaa gtgacgggcc
      661 gagcaaggga tctagagtcg actccggaat aacttcgtat aggatacttt atacgaagtt
      721 atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc
      781 gtatagcata cattatacga agttatggtt agacgttgat cctggcgctg gcgcaagcag
      841 cagggtgtct atccatgccg ctctcctggg cgcagctcat gggcagggtg ccggcggcct
      901 gctcctccac ctcgggaggg aagccgtggc tgttaaggtg gatcacaaag tggcttaagc
      961 tactgaacca gttcttgcaa gcagaattca cggcgatctt gccgcccttc ttggccttaa
     1021 tgagaatctc gcggatcttg cgggcgtcca acttgccggt cagtccttta ggcacctcgt
     1081 ccacgaacac aacaccaccg cgcagcttct tggcggttgt aacctggctg gccacatagt
     1141 ccacgatctc cttctcggtc atggttttac cgtgttccag cacgacgact gcggcgggca
     1201 gctcgccggc atcgtcgtcg ggcaggccgg cgaccccggc gtcgaagatg ttggggtgtt
     1261 gcagcaggat gctctccagt tcggctgggg ctacctggta gcccttgtat ttgatcaggc
     1321 tcttcagccg gtccacgatg aagaagtgct cgtcctcgtc ccagtaggcg atgtcgccgc
     1381 tgtgcagcca gccgtccttg tcgatgagag cgtttgtagc ctcggggttg ttaacgtagc
     1441 cgctcatgat catggggcca cggacgcaca gctcgccgcg ctggttcaca cccagtgtct
     1501 taccggtgtc caagtccacc accttagcct cgaagaaggg caccaccttg cctactgcgc
     1561 caggcttgtc gtccccttcg ggggtgatca gaatggcgct ggttgtttct gtcaggccgt
     1621 agccctggcg gatgcctggt aggtggaagc gtttggccac ggcctcacct acctccttgc
     1681 tgagcggcgc cccgccgctg gcgatctcgt gcaagttgct taggtcgtac ttgtcgatga
     1741 gagtgctctt agcgaagaag ctaaatagtg tgggcaccag cagggcagat tgaatcttat
     1801 agtcttgcaa gctgcgcaag aatagctcct cctcgaagcg gtacatgagc acgacccgaa
     1861 agccgcagat caagtagccc agcgtggtga acatgccgaa gccgtggtga aatggcacca
     1921 cgctgaggat agcggtgtcg gggatgatct ggttgccgaa gatggggtcg cgggcatgac
     1981 tgaatcggac acaagcggtg cggtgcggta gggctacgcc cttgggcaat ccggtactgc
     2041 cactactgtt catgatcagg gcgatggttt tgtcccggtc gaagctctcg ggcacgaagt
     2101 cgtactcgtt gaagccgggt ggcaaatggg aagtcacgaa ggtgtacatg ctttggaagc
     2161 cctggtagtc ggtcttgcta tccatgatga tgatcttttg tatgatcggt agcttctttt
     2221 gcacgttgag gatcttttgc agccctttct tgctcacgaa tacgacggtg ggctggctga
     2281 tgcccatgct gttcagcagc tcgcgctcgt tgtagatgtc gttagctggg gccacagcca
     2341 caccgatgaa cagggcaccc aacacgggca tgaagaactg caagctattc tcgctgcaca
     2401 ccacgatccg atggtttgta ttcagcccat agcgcttcat agcttctgcc agccgaacgc
     2461 tcatctcgaa gtactcggcg taggtaatgt ccacctcgat atgtgcgtcg gtaaaggcga
     2521 tggtgccggg caccagggcg tagcgcttca tggctttgtg cagctgctcg ccggcggtcc
     2581 cgtcttcgag tgggtagaat ggcgctgggc ccttcttaat gtttttggca tcttcacctg
     2641 cagggccctg aaaatacagg ttttcggatc ctgaaccgcc tgccgatccc tcatgccact
     2701 ctattttctg agcttcgaaa atatcgttca ggccgctagc tgcggccgct ccacccttat
     2761 cgtcatcgtc cttgtagtcg atgtcatgat ctttataatc accgtcatgg tctttgtagt
     2821 cctgcaaagg aagatagata gataagatct ccacacgcct cttcccccaa ccagccgata
     2881 aaactgacca gggacatcac accagagatt ttaaagaaaa tattaccgta tgctgggtgg
     2941 tggctttggg acaaaggact tgcctggagt gcctgagacc catagtatgg cctccatact
     3001 tcccctcaaa ataaaagtca gagctggaga atcagctcaa tggtgaagag aagcacccac
     3061 acggctgctt acacccatct gtaactcaca gttccagggg atccaatgcc ctcttctgtc
     3121 ccctatggtc accaaacatg caagtggtgc accgacatac atacaggcaa aacacccata
     3181 tactttaaag taaaaattta aaaaaggaaa agtcatgtta aaactgaaac ataagagctt
     3241 cagctattag gagacttgaa cactgagaga cagaaatgaa aaggaatatt ctgtgatcga
     3301 atgcatgtgt gtgagcaggc tcctccgagg tcttactgga ctgtctgtta agtgcatcag
     3361 atacaaaaat gcaaagatga cgtggtatcg ttatttggaa ggacaaagta caataaatac
     3421 catctacagg attaagaagt actgccatcc tgcaagtcct ggtacttgct gcttgaagac
     3481 gggcgacaaa ctatggaggg agccacccgt gtcctgtgtg cgagggcctg gcctgaggct
     3541 gacctacctt gtcatcgtca tccttgtagt ccatggtggc ctctccttcg agtgcaaaaa
     3601 gaacaagtag cttgtattct atagtgtcac ctaaatgcta gcctatagtg agtcgtatta
     3661 agtactctag ccttaagagc tgtaattgaa ctgggagtgg acacctgtgg agagaaaggc
     3721 aaagtggatg tcagtaagac caataggtgc ctatcagaaa cgcaagagtc ttctctgtct
     3781 cgacaagccc agtttctatt ggtctcctta aacctgtctt gtaaccttga tacttacctg
     3841 cccagtgcct cacgaccaac tttcgctacc ttaggaccgt tatagttact gcagcgctag
     3901 cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact
     3961 gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg
     4021 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     4081 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4141 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4201 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4261 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4321 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4381 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4441 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     4501 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4561 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4621 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     4681 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     4741 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     4801 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     4861 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     4921 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     4981 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     5041 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     5101 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     5161 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     5221 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     5281 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     5341 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     5401 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5461 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5521 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5581 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5641 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5701 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     5761 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5821 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5881 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5941 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     6001 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     6061 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6121 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6181 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6241 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6301 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6361 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6421 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6481 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6541 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6601 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6661 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6721 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6781 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6841 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6901 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6961 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7021 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7081 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7141 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7201 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7261 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7321 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7381 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7441 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7501 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7561 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7621 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7681 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7741 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7801 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7861 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
