LOCUS       sequence-207058-        6454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Per2 transcription fluorescence reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6454)
  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 6454)
  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-mCherry-Venus-NLS-D2
FEATURES             Location/Qualifiers
     source          1..6454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    complement(700..733)
                     /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(784..817)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      826..835
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             832..1542
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(977..995)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1245..1264
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(1691..1711)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(1715..2425,2426..2428,2429..2431))
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1757..1778)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2106..2125
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2365..2386
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2428..2437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2439..2472
                     /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    2523..2556
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2581..3169
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2634..2654)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3052..3063)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3201..3677
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3400..3419)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3717..3857
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3717..3846
                     /label=AAV2 ITR
     rep_origin      3932..4387
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4019..4038)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4229..4250
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4404..4423)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4523..4545
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4583..4601)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4669..4773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4774..4842,4843..5634)
                     /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(4992..5011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5805..6393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6294..6313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtcggat taccgaggct ggtcacgtcg
      181 tcgcaggtga taggccggtg gccctggtct ctgccggctg tgagttgcgc agcggccaag
      241 caccattccc ccgcgccgca gtggtacgcg ccactccggg gctgcacgag cgggccaccg
      301 ccgtgccagg tgaatggaag tcccgcaggc cggaagtgga cgagcctact cgcccgggcg
      361 cgggggggcg caagagcgcg cagcatcttc attgaggaac ccgggcggcg aacatggagt
      421 tccatgtgcg tcttatgtaa agagagcgac gggcgtctcc accaattgat gagcgtagct
      481 ctcaggttcc gccccgccag tatgcaaatg aggtggcact ccgaccaatg gcgcgcgcag
      541 gggcgggctc agcgcgcgcg gtcacgtttt ccactatgtg acagcggagg gcgacgcggc
      601 ggcagcggcg ctactgggac tagcggctcc gggcggctgc ggcgcaggcc gagcgcacca
      661 agtgacgggc cgagcaaggg atctagagtc gactccggaa taacttcgta taggatactt
      721 tatacgaagt tatgcagaat ggtagctgga ttgtagctgc tattagcaat atgaaacctc
      781 ttaataactt cgtatagcat acattatacg aagttatggc gcgccgccac catggtgagc
      841 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
      901 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      961 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     1021 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     1081 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1141 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     1201 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     1261 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     1321 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     1381 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     1441 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     1501 ggccgccact ccaccggcgg catggacgag ctgtacaagt aggctacaca ttgatcctag
     1561 cagaagcaca ggctgcaggg tgacggtcca tcccgctctc ctgggcacaa gacatgggca
     1621 gcgtgccatc atcctgctcc tccacctccg gcgggaagcc atggctaagc ttagaattca
     1681 cggggtcttc tacctttctc ttcttttttg gaggcttgta cagctcgtcc atgccgagag
     1741 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     1801 ctttgctcag tttggactgg tagctcaggt agtggttgtc gggcagcagc acggggccgt
     1861 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgccgccg tcctcgatgt
     1921 tgtggcggat cttgaagttt gccttgatgc cgttcttctg cttgtcggcg gtgatataga
     1981 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     2041 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     2101 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     2161 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc
     2221 actgcaggcc gtagccgagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     2281 tggtgcagat caacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     2341 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     2401 cggtgaacag ctcctcgccc ttgctcacca tggtggccat aacttcgtat aaagtatcct
     2461 atacgaagtt atttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa
     2521 gaataacttc gtataatgta tgctatacga agttatgaat tcgatatcaa gcttatcgat
     2581 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2641 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2701 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2761 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2821 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2881 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2941 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3001 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3061 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3121 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     3181 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     3241 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     3301 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     3361 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     3421 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     3481 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     3541 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     3601 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     3661 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     3721 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3781 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3841 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3901 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3961 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4021 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4081 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4141 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4201 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4261 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4321 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4381 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4441 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4501 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4561 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4621 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4681 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4741 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4801 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4861 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4921 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4981 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5041 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5101 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5161 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5221 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5281 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5341 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5401 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5461 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5521 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5581 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5641 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5701 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5761 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5821 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5881 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5941 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6001 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6061 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6121 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6181 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6241 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6301 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6361 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6421 ttcctggcct tttgctggcc ttttgctcac atgt
//
