LOCUS       sequence-176458-        7793 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Localizes CRY2 to excitatory synapses.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7793)
  AUTHORS   Sinnen BL, Bowen AB, Forte JS, Hiester BG, Crosby KC, Gibson ES, 
            Dell'Acqua ML, Kennedy MJ
  TITLE     Optogenetic Control of Synaptic Composition and Function.
  JOURNAL   Neuron. 2017 Feb 8;93(3):646-660.e5. doi: 
            10.1016/j.neuron.2016.12.037. Epub 2017 Jan 26.
  PUBMED    28132827
REFERENCE   2  (bases 1 to 7793)
  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"; doi: 
            "10.1016/j.neuron.2016.12.037"; journalName: "Neuron"; date: 
            "2017-02-8- 8"; volume: "93"; issue: "3"; pages: "646-660.e5"
COMMENT     Sequence Label: CRY2-GFP-homer1c
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     319..341
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     403..422
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(462..2081,2082..2087,2088..2297)
                     /codon_start=1
                     /product="Arabidopsis thaliana cryptochrome 2 in which the 
                     nuclear localization signal was removed by mutating 
                     residues 541 and 542 to alanine"
                     /label=CRY2(Delta-NLS)
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAAPDEIVADSFEALGANTIKE
                     PGLCPSVSSNDQQVPSAVRYNGSAAVKPEEEEERDMKKSRGFDERELFSTAESSSSSSV
                     FFVSQSCSLASEGKNLEGIQDSSDQITTSLGKNGCK"
     regulatory      2316..2325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2322..2324,2325..2327,2328..3038)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2322..2324,2325..2327,2328..3038)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2367..2388)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2628..2647)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2975..2996
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4283..4302)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4348..4403
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4402..4421)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4794..4810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4794..4810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4807..4829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4818..4834
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4842..4848,4849..4866,4867..4872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4887..4908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4967..5162
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5013..5148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5168..5302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5205..5224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5259..5278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5370..5387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5541..6129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5621..5640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6300..7091,7092..7160))
                     /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     6923..6942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7161..7265)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7296..7675
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
       61 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
      121 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
      181 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
      241 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
      301 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
      361 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
      421 tgctgtctca tcattttggc aaagaattgc tcgaggccac catgaagatg gacaaaaaga
      481 ctatagtttg gtttagaaga gacctaagga ttgaggataa tcctgcatta gcagcagctg
      541 ctcacgaagg atctgttttt cctgtcttca tttggtgtcc tgaagaagaa ggacagtttt
      601 atcctggaag agcttcaaga tggtggatga aacaatcact tgctcactta tctcaatcct
      661 tgaaggctct tggatctgac ctcactttaa tcaaaaccca caacacgatt tcagcgatct
      721 tggattgtat ccgcgttacc ggtgctacaa aagtcgtctt taaccacctc tatgatcctg
      781 tttcgttagt tcgggaccat accgtaaagg agaagctggt ggaacgtggg atctctgtgc
      841 aaagctacaa tggagatcta ttgtatgaac cgtgggagat atactgcgaa aagggcaaac
      901 cttttacgag tttcaattct tactggaaga aatgcttaga tatgtcgatt gaatccgtta
      961 tgcttcctcc tccttggcgg ttgatgccaa taactgcagc ggctgaagcg atttgggcgt
     1021 gttcgattga agaactaggg ctggagaatg aggccgagaa accgagcaat gcgttgttaa
     1081 ctagagcttg gtctccagga tggagcaatg ctgataagtt actaaatgag ttcatcgaga
     1141 agcagttgat agattatgca aagaacagca agaaagttgt tgggaattct acttcactac
     1201 tttctccgta tctccatttc ggggaaataa gcgtcagaca cgttttccag tgtgcccgga
     1261 tgaaacaaat tatatgggca agagataaga acagtgaagg agaagaaagt gcagatcttt
     1321 ttcttagggg aatcggttta agagagtatt ctcggtatat atgtttcaac ttcccgttta
     1381 ctcacgagca atcgttgttg agtcatcttc ggtttttccc ttgggatgct gatgttgata
     1441 agttcaaggc ctggagacaa ggcaggaccg gttatccgtt ggtggatgcc ggaatgagag
     1501 agctttgggc taccggatgg atgcataaca gaataagagt gattgtttca agctttgctg
     1561 tgaagtttct tctccttcca tggaaatggg gaatgaagta tttctgggat acacttttgg
     1621 atgctgattt ggaatgtgac atccttggct ggcagtatat ctctgggagt atccccgatg
     1681 gccacgagct tgatcgcttg gacaatcccg cgttacaagg cgccaaatat gacccagaag
     1741 gtgagtacat aaggcaatgg cttcccgagc ttgcgagatt gccaactgaa tggatccatc
     1801 atccatggga cgctccttta accgtactca aagcttctgg tgtggaactc ggaacaaact
     1861 atgcgaaacc cattgtagac atcgacacag ctcgtgagct actagctaaa gctatttcaa
     1921 gaacccgtga agcacagatc atgatcggag cagcacctga tgagattgta gcagatagct
     1981 tcgaggcctt aggggctaat accattaaag aacctggtct ttgcccatct gtgtcttcta
     2041 atgaccaaca agtaccttcg gctgttcgtt acaacgggtc agcggcagtg aaacctgagg
     2101 aagaagaaga gagagacatg aagaaatcta ggggattcga tgaaagggag ttgttttcga
     2161 ctgctgaatc ttcttcttct tcgagtgtgt ttttcgtttc gcagtcttgc tcgttggcat
     2221 cagaagggaa gaatctggaa ggtattcaag attcatctga tcagattact acaagtttgg
     2281 gaaaaaatgg ttgcaaagcc cgggatccac cggtcgccac catggtgagc aagggcgagg
     2341 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2401 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2461 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2521 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2581 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2641 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2701 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2761 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2821 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2881 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2941 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3001 ccgccgggat cactctcggc atggacgagc tgtacaagga attcggtggt ggtggtgggt
     3061 cgaccatggg ggaacaacct atcttcagca ctcgagctca tgtcttccag atcgacccaa
     3121 acacaaagaa gaactgggta cccaccagca agcatgcagt tactgtgtct tatttctatg
     3181 acagcacaag gaatgtgtat aggataatca gtttagacgg ctcaaaggca ataataaata
     3241 gcaccatcac tccaaacatg acatttacta aaacatctca aaagtttggc caatgggctg
     3301 atagccgggc aaacactgtt tatggactgg gattctcctc tgagcatcat ctctcaaaat
     3361 ttgcagaaaa gtttcaggaa tttaaagaag ctgctcggct ggcaaaggag aagtcgcagg
     3421 agaagatgga actgaccagt accccttcac aggaatcagc aggaggagat cttcagtctc
     3481 ctttaacacc agaaagtatc aatgggacag atgatgagag aacacccgat gtgacacaga
     3541 actcagagcc aagggctgag ccagctcaga atgcattgcc attttcacat agtgccgggg
     3601 atcgaaccca gggcctctct catgctagtt cagccatcag caaacactgg gaggctgaac
     3661 tagccacgct caaggggaac aatgccaagc tcaccgcagc gctgctggag tccactgcca
     3721 acgtgaagca gtggaagcaa cagctggctg cctaccagga ggaggcagag cggctgcaca
     3781 agcgggtcac ggagctggaa tgtgttagta gtcaagcaaa cgcggtgcac agccacaaga
     3841 cagagctgag tcagacagtg caggagctgg aagagaccct aaaagtaaag gaagaggaaa
     3901 tagaaagatt aaaacaagaa attgataacg ccagagaact tcaagaacag agggactctt
     3961 tgactcagaa actacaggaa gttgagattc gaaataaaga cctggagggg cagctgtcgg
     4021 agctggagca gcgcctggag aagagccaga gcgagcagga cgctttccgc agtaacctga
     4081 agactctcct agagattctg gacgggaaaa tatttgaact aacagaattg cgggataatt
     4141 tggccaagct actagaatgc agctaagaat tctgcagata tccatcacac tggcggccgc
     4201 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     4261 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     4321 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     4381 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     4441 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     4501 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     4561 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     4621 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     4681 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
     4741 accgtcgacc agctggtcga cggtgcaccg tcgaccagct tggcgtaatc atggtcatag
     4801 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4861 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4921 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     4981 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5041 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5101 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5161 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5221 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5281 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     5341 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5941 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6121 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6241 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6301 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6361 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6421 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6481 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6541 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6601 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6661 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6721 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6781 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6841 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6901 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6961 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7021 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7081 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7141 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7261 ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga ctagttatta
     7321 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7381 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7441 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7501 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7561 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7621 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     7681 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     7741 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggg
//
