LOCUS       sequence-210742-        7676 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 7676)
  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 7676)
  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)-forward-intron336-Venus-NLS-D2
FEATURES             Location/Qualifiers
     source          1..7676
                     /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(1721..1754)
                     /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(1805..1838)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          1898..2030
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2013..2032
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        2075..2093
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2099..2117
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2099..2116
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      2155..2164
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2164..2187
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2578..2643
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2665..2709
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2734..2754
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2764..2766,2767..3477)
                     /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(2806..2827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3067..3086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3414..3435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3481..3501
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    3661..3694
                     /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    3745..3778
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3803..4391
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3856..3876)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4274..4285)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4423..4899
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4622..4641)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4939..5079
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4939..5068
                     /label=AAV2 ITR
     rep_origin      5154..5609
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5241..5260)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5451..5472
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5626..5645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5745..5767
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5805..5823)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5891..5995
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5996..6064,6065..6856)
                     /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(6214..6233)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7027..7615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7516..7535
                     /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 cgtgtaaaga tgcacatgtg gccctgtgtt ctcattagac
      181 ttcaattgct ttgataaaac atcatgagca aaagcaactt ggccaggaag gaaggggttt
      241 gttgccgttt acagttaata atccatcgtg gggagaagtc agggcagaaa ttcaaagcag
      301 aaacctggag atagagggct gaagcagaag ccatggagga aagccgtttt ggttccctgt
      361 ggcttcctca gcctgcttcc atatacaacc cgcctagggc taccagccta atggtggcgc
      421 cacacacact ggtcgggtcc ttccacatca ctcattaatc aagaaagtgc ccagacttcc
      481 ctacaggcaa cctgatggaa gtctttacta caattaaggt tcctcttccc atttaacttc
      541 agttgaagaa aaaaaaaaaa aaaaaaaaaa aaagcaaagc aaaaagccca aaaacctaat
      601 caaaatacct tctaaactac acttcatgta atagattaga gttgataaat tgacgatgta
      661 tctacagtat tatgaaatgc tgaatatctg agatcggacc tgtcttctcc agagcttggg
      721 caaaggtgcc cgccccaaac agtaaaaaag gctaacggcg tgatttctgc tcctttcaga
      781 gcaggggact ccgattaaca cctgattgaa gctgggtgag aggcaggact gtgcctcggt
      841 tgcctctcac ccagcccagc aataactctc agggcacacg gaactggctt cagaccttcc
      901 acacaaccca taggaagagc cctgggcgcg ctcccgcgtc ctagcctctt ctcttccacc
      961 gcgagagcgc agcgcacgcg cgccgtgccc ggcagagcgc gcgcccgcga gctccccgca
     1021 gccccgtgct catccgggtc cccgcgagcc tcggccaatg gaggcggctc ccggccccgg
     1081 gagcagaact atgcctcctc ccccttctcg cggcctgtgg cccccgcgag cggcctgggg
     1141 ctcaggagac ccgcagaggc tgcgagtccg gatcccacgc gagaactcag gtcgtgagcg
     1201 cccagcgatc gcaaccggtc ccgaagcacc gggcgcggcg gcgcacgcgg gggtctgagc
     1261 cagtgtagta aacacacttc agaaacgtga ggtgccggtg gtcacgtggg gtgcatgccg
     1321 cccaatgaga agccgggggc ggggcggagg ccgctgacgc ggcggagtca cccgggcagc
     1381 ctcgggaccg gtgaccggca ggcgagcgtc ccgcggcctg gagctctagc ccgcgctctc
     1441 cgagctcgct gcccactccg ggacgcacag gccgcgtcgc gcggaggccg cgtgagctcg
     1501 tgtccgttcg tggaggccgc ctcgggcgcg ccaccgtctt ctcgcgggtc gcctcggttg
     1561 ggcgaggttt ccgtgacgag cctcccgggg ctgctgcgac cagcccgggc cgtcgtggcg
     1621 gctcgggccc cgcgcgctcg cttgcgctcc gggggtctca ggccctggaa ggtatgcgtg
     1681 tcctccggac gagatctgcg atctaagtgt cgactccgga ataacttcgt ataggatact
     1741 ttatacgaag ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct
     1801 cttaataact tcgtatagca tacattatac gaagttatgg ctgcagtaac tataacggtc
     1861 ctaaggtagc gaaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
     1921 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
     1981 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
     2041 ccagttcaat tacagctctt aaggctagag tacttaatac gactcactat aggctagcat
     2101 ttaggtgaca ctatagaata caagctactt gttctttttg cactcgaagg agaggccacc
     2161 atggactaca aggatgacga tgacaagaat taccctgtta tccctaatcg agatggttgt
     2221 tatgacacag tgtagaaact atggcatagg acagatgact gtgttatatg aattcataat
     2281 gtcttgtttt cagtagcagt gggattatgt tgtatcttgc cattgccttt ttcagactat
     2341 tttatagaca gaactagaaa tactagttat ataagctgtc ctagcacaga ctagaaagta
     2401 ggtcattgtg atgggagtat gctaaaccac ccactggttg ctatagcgat gacctacttt
     2461 agaaactatt tctgttgatt catgttgaaa tcatcttttg gaagtggcat aaggaagtta
     2521 ctacatgtac gtttttatag tagtaaaagg tagagctgtc aagaggtacc caccatggac
     2581 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga cgatgacgat
     2641 aagggtggag cggccgcagc tagcggcctg aacgatattt tcgaagctca gaaaatagag
     2701 tggcatgagg gatcggcagg cggttcagga tccgaaaacc tgtattttca gggccctgca
     2761 ggtgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2821 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2881 ggcaagctga ccctgaagtt gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2941 ctcgtgacca ccctcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
     3001 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3061 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3121 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3181 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
     3241 ggcatcaagg caaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     3301 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3361 tacctgagct accagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3421 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagcct
     3481 ccaaaaaaga agagaaaggt agaagacccc gtgaattcta agcttagcca tggcttcccg
     3541 ccggaggtgg aggagcagga tgatggcacg ctgcccatgt cttgtgccca ggagagcggg
     3601 atggaccgtc accctgcagc ctgtgcttct gctaggatca atgtgtaggg atccgctagc
     3661 ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg
     3721 ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga
     3781 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3841 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3901 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3961 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4021 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4081 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4141 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4201 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     4261 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4321 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4381 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     4441 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     4501 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     4561 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     4621 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     4681 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     4741 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     4801 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     4861 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4921 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4981 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     5041 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     5101 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     5161 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5221 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5281 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5341 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     5401 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5461 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     5521 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5581 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5641 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5701 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5761 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5821 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5881 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5941 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6001 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6061 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6121 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6181 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6241 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6301 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6361 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6421 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6481 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6541 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6601 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6661 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6721 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6781 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6841 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6901 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6961 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7021 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7081 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7141 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7201 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7261 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7321 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7381 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7441 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7501 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7561 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7621 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
