LOCUS       sequence-210760-        7672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cry1 transcription fluorescence reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7672)
  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 7672)
  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(Cry1)-DIO-intron336-Venus-NLS-D2
FEATURES             Location/Qualifiers
     source          1..7672
                     /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(1722..1755)
                     /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(1806..1839)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1989..2009)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(2013..2723,2724..2726))
                     /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="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2055..2076)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2404..2423
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2663..2684
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2736..2756)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2781..2825)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(2847..2912)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3303..3326)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3326..3335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3373..3391)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3374..3391)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(3397..3415)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3458..3477)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3460..3592
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    3657..3690
                     /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    3741..3774
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3799..4387
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3852..3872)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4270..4281)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4419..4895
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4618..4637)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4935..5075
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4935..5064
                     /label=AAV2 ITR
     rep_origin      5150..5605
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5237..5256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5447..5468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5622..5641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5741..5763
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5801..5819)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5887..5991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5992..6060,6061..6852)
                     /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(6210..6229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7023..7611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7512..7531
                     /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 gcgtgtaaag atgcacatgt ggccctgtgt tctcattaga
      181 cttcaattgc tttgataaaa catcatgagc aaaagcaact tggccaggaa ggaaggggtt
      241 tgttgccgtt tacagttaat aatccatcgt ggggagaagt cagggcagaa attcaaagca
      301 gaaacctgga gatagagggc tgaagcagaa gccatggagg aaagccgttt tggttccctg
      361 tggcttcctc agcctgcttc catatacaac ccgcctaggg ctaccagcct aatggtggcg
      421 ccacacacac tggtcgggtc cttccacatc actcattaat caagaaagtg cccagacttc
      481 cctacaggca acctgatgga agtctttact acaattaagg ttcctcttcc catttaactt
      541 cagttgaaga aaaaaaaaaa aaaaaaaaaa aaaagcaaag caaaaagccc aaaaacctaa
      601 tcaaaatacc ttctaaacta cacttcatgt aatagattag agttgataaa ttgacgatgt
      661 atctacagta ttatgaaatg ctgaatatct gagatcggac ctgtcttctc cagagcttgg
      721 gcaaaggtgc ccgccccaaa cagtaaaaaa ggctaacggc gtgatttctg ctcctttcag
      781 agcaggggac tccgattaac acctgattga agctgggtga gaggcaggac tgtgcctcgg
      841 ttgcctctca cccagcccag caataactct cagggcacac ggaactggct tcagaccttc
      901 cacacaaccc ataggaagag ccctgggcgc gctcccgcgt cctagcctct tctcttccac
      961 cgcgagagcg cagcgcacgc gcgccgtgcc cggcagagcg cgcgcccgcg agctccccgc
     1021 agccccgtgc tcatccgggt ccccgcgagc ctcggccaat ggaggcggct cccggccccg
     1081 ggagcagaac tatgcctcct cccccttctc gcggcctgtg gcccccgcga gcggcctggg
     1141 gctcaggaga cccgcagagg ctgcgagtcc ggatcccacg cgagaactca ggtcgtgagc
     1201 gcccagcgat cgcaaccggt cccgaagcac cgggcgcggc ggcgcacgcg ggggtctgag
     1261 ccagtgtagt aaacacactt cagaaacgtg aggtgccggt ggtcacgtgg ggtgcatgcc
     1321 gcccaatgag aagccggggg cggggcggag gccgctgacg cggcggagtc acccgggcag
     1381 cctcgggacc ggtgaccggc aggcgagcgt cccgcggcct ggagctctag cccgcgctct
     1441 ccgagctcgc tgcccactcc gggacgcaca ggccgcgtcg cgcggaggcc gcgtgagctc
     1501 gtgtccgttc gtggaggccg cctcgggcgc gccaccgtct tctcgcgggt cgcctcggtt
     1561 gggcgaggtt tccgtgacga gcctcccggg gctgctgcga ccagcccggg ccgtcgtggc
     1621 ggctcgggcc ccgcgcgctc gcttgcgctc cgggggtctc aggccctgga aggtatgcgt
     1681 gtcctccgga cgagatctgc gatctaagtg tcgactccgg aataacttcg tataggatac
     1741 tttatacgaa gttatgcaga atggtagctg gattgtagct gctattagca atatgaaacc
     1801 tcttaataac ttcgtatagc atacattata cgaagttatg gctacacatt gatcctagca
     1861 gaagcacagg ctgcagggtg acggtccatc ccgctctcct gggcacaaga catgggcagc
     1921 gtgccatcat cctgctcctc cacctccggc gggaagccat ggctaagctt agaattcacg
     1981 gggtcttcta cctttctctt cttttttgga ggcttgtaca gctcgtccat gccgagagtg
     2041 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     2101 ttgctcagtt tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     2161 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg
     2221 tggcggatct tgaagtttgc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
     2281 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     2341 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     2401 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     2461 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     2521 tgcaggccgt agccgagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     2581 gtgcagatca acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     2641 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     2701 gtgaacagct cctcgccctt gctcacacct gcagggccct gaaaatacag gttttcggat
     2761 cctgaaccgc ctgccgatcc ctcatgccac tctattttct gagcttcgaa aatatcgttc
     2821 aggccgctag ctgcggccgc tccaccctta tcgtcatcgt ccttgtagtc gatgtcatga
     2881 tctttataat caccgtcatg gtctttgtag tccatggtgg gtacctcttg acagctctac
     2941 cttttactac tataaaaacg tacatgtagt aacttcctta tgccacttcc aaaagatgat
     3001 ttcaacatga atcaacagaa atagtttcta aagtaggtca tcgctatagc aaccagtggg
     3061 tggtttagca tactcccatc acaatgacct actttctagt ctgtgctagg acagcttata
     3121 taactagtat ttctagttct gtctataaaa tagtctgaaa aaggcaatgg caagatacaa
     3181 cataatccca ctgctactga aaacaagaca ttatgaattc atataacaca gtcatctgtc
     3241 ctatgccata gtttctacac tgtgtcataa caaccatctc gattagggat aacagggtaa
     3301 ttcttgtcat cgtcatcctt gtagtccatg gtggcctctc cttcgagtgc aaaaagaaca
     3361 agtagcttgt attctatagt gtcacctaaa tgctagccta tagtgagtcg tattaagtac
     3421 tctagcctta agagctgtaa ttgaactggg agtggacacc tgtggagaga aaggcaaagt
     3481 ggatgtcagt aagaccaata ggtgcctatc agaaacgcaa gagtcttctc tgtctcgaca
     3541 agcccagttt ctattggtct ccttaaacct gtcttgtaac cttgatactt acctgcccag
     3601 tgcctcacga ccaactttcg ctaccttagg accgttatag ttactgcagc gctagcataa
     3661 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat
     3721 tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc
     3781 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3841 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3901 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3961 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4021 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4081 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4141 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     4201 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     4261 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4321 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4381 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     4441 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     4501 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     4561 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     4621 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     4681 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     4741 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     4801 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     4861 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     4921 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4981 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     5041 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     5101 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     5161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5221 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5341 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     5401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5461 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     5521 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5581 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     5641 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5701 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5761 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5821 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
