LOCUS       sequence-383962-        7377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  all-optical silencing method.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7377)
  AUTHORS   Fan LZ, Kim DK, Jennings JH, Tian H, Wang PY, Ramakrishnan C, 
            Randles S, Sun Y, Thadhani E, Kim YS, Quirin S, Giocomo L, Cohen AE,
            Deisseroth K
  TITLE     All-optical physiology resolves a synaptic basis for behavioral 
            timescale plasticity.
  JOURNAL   Cell. 2023 Feb 2;186(3):543-559.e19. doi: 
            10.1016/j.cell.2022.12.035. Epub 2023 Jan 19.
  PUBMED    36669484
REFERENCE   2  (bases 1 to 7377)
  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.cell.2022.12.035"; journalName: "Cell"; date: "2023-02-2-
            2"; volume: "186"; issue: "3"; pages: "543-559.e19"
COMMENT     Sequence Label: pAAV-CaMKIIa-DIO eHcKCR1 3.0-oScarlett-KV2.1
FEATURES             Location/Qualifiers
     source          1..7377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        138..1426
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     protein_bind    complement(1447..1480)
                     /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(1531..1564)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1795..2487)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWDRVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2070..2089)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2339..2357
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    3362..3395
                     /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    3446..3479
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3504..4092
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3557..3577)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3975..3986)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4124..4600
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4323..4342)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4640..4780
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4640..4769
                     /label=AAV2 ITR
     rep_origin      4855..5310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4942..4961)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5152..5173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5327..5346)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5446..5468
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5506..5524)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5592..5696
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5697..5765,5766..6557)
                     /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(5915..5934)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6728..7316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7217..7236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgg gcgacctttg gtcgcccggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      121 cggccgcacg cgtttaacat tatggcctta ggtcacttca tctccatggg gttcttcttc
      181 tgattttcta gaaaatgaga tgggggtgca gagagcttcc tcagtgacct gcccagggtc
      241 acatcagaaa tgtcagagct agaacttgaa ctcagattac taatcttaaa ttccatgcct
      301 tgggggcatg caagtacgat atacagaagg agtgaactca ttagggcaga tgaccaatga
      361 gtttaggaaa gaagagtcca gggcagggta catctacacc acccgcccag ccctgggtga
      421 gtccagccac gttcacctca ttatagttgc ctctctccag tcctaccttg acgggaagca
      481 caagcagaaa ctgggacagg agccccagga gaccaaatct tcatggtccc tctgggagga
      541 tgggtgggga gagctgtggc agaggcctca ggaggggccc tgctgctcag tggtgacaga
      601 taggggtgag aaagcagaca gagtcattcc gtcagcattc tgggtctgtt tggtacttct
      661 tctcacgcta aggtggcggt gtgatatgca caatggctaa aaagcaggga gagctggaaa
      721 gaaacaagga cagagacaga ggccaagtca accagaccaa ttcccagagg aagcaaagaa
      781 accattacag agactacaag ggggaaggga aggagagatg aattagcttc ccctgtaaac
      841 cttagaaccc agctgttgcc agggcaacgg ggcaatacct gtctcttcag aggagatgaa
      901 gttgccaggg taactacatc ctgtctttct caaggaccat cccagaatgt ggcacccact
      961 agccgttacc atagcaactg cctctttgcc ccacttaatc ccatcccgtc tgttaaaagg
     1021 gccctatagt tggaggtggg ggaggtagga agagcgatga tcacttgtgg actaagtttg
     1081 ttcgcatccc cttctccaac cccctcagta catcaccctg ggggaacagg gtccacttgc
     1141 tcctgggccc acacagtcct gcagtattgt gtatataagg ccagggcaaa gaggagcagg
     1201 ttttaaagtg aaaggcaggc aggtgttggg gaggcagtta ccggggcaac gggaacaggg
     1261 cgtttcggag gtggttgcca tggggacctg gatgctgacg aaggctcgcg aggctgtgag
     1321 cagccacagt gccctgctca gaagccccaa gctcgtcagt caagccggtt ctccgtttgc
     1381 actcaggagc acgggcaggc gagtggcccc tagttctggg ggcagctcta gagcggtacc
     1441 ggatccataa cttcgtatag gatactttat acgaagttat gcagaatggt agctggattg
     1501 tagctgctat tagcaatatg aaacctctta ataacttcgt atagcataca ttatacgaag
     1561 ttatggcgcg ccctattaca cctcgttctc gtagcagaag aatctggtgg cttcagggaa
     1621 gtccgtggca cagctgatga agctgtcaat gctggacatg ctccgcatgt cgatgacgcc
     1681 ctccgtgcgt gcgggcagag gggccacggg gctgggcatg ctgctcatct ccagctcctc
     1741 tggaggcttg ctctgcgggg ccatctcctt ggtgttgagg atgggctggg actgcttgta
     1801 cagctcgtcc atgccgccgg tggagtggcg gccctcggag cgttcgtact gttccaccac
     1861 ggtgtagtcc tcgttgtggg aggtgatgtc caacttgcgg tccacgttgt aggcgccggg
     1921 catctgcacg ggcttcttgg ccttgtaggt ggtcttgaag tccgccaggt agcgcccgcc
     1981 gtccttcagg cgcagggcca tcttaatgtc gcccttcagc acgccgtcct cggggtacaa
     2041 ccgctcggtg gacgcttccc agcccattgt cttcttctgc attacggggc cgtcaggagg
     2101 gaagttggtg ccgcggagct tcaccttgta gatcagggtg ccgtcctcca gggaggtgtc
     2161 ctgggtcacg gtcacggcgc cgccgtcctc gaagttcatc acgcggtccc acttgaagcc
     2221 ctcggggaag gactgcttat agtagtcggg gatgtcggcg gggtgcttgg tgaaggccct
     2281 ggagccgtac atgaactgag gggacaggat gtcccaggag aagggcaggg ggccaccctt
     2341 ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc
     2401 gatctcgaac tcgtggccgt tcatggagcc ctccatgtgc accttgaacc gcatgaactc
     2461 cttgatcact gcctcgccct tgctcaccac gttgatgtcg atctggtcca gggggatgta
     2521 ctcgccctcg ctggtgatcc tgctcttggc ggccgcggag agtgtctcag gctcattcag
     2581 gggcaggccc tgcacctcgg tgtgctcttc cagttccagt ctaaacacgg tcatcaggat
     2641 gccgaagatg cttttacagg tcaggtccag gaaggcgtgg atgatgtaaa aggcctgttg
     2701 gctgatgtgg cctgtgccca gccagctgaa gatgaagagg atggggaaca taggccagaa
     2761 gccaaaaaag atgacgcatg cccagaacag ccaccgctgg cacttaggag ggtacaccag
     2821 gtatctttcc ttcacgtgcc gcatcagaga cagggcgaag ggggtgaacc agcaacaacc
     2881 cagggcgtaa taagcgaacc tgtaggcgct aggggtcaca aaagcagcaa cgccgcacca
     2941 cagtgtcagg aacacggcga agaagtatct gatcttatga ggcagatcca gtgtgaaggc
     3001 gaaatccagg atcagcagag ggcaggtgaa cacgtaatcc agataccagt accagttcag
     3061 ggaaaatggg gttcctccgc cgtcgaagaa gtcgaaggac ttggcggcgt acatggtgcc
     3121 acaaagcagg cagatgatgc cgctcaggct gttcacctca ggggctctgc cgctcttgta
     3181 aggtttacaa gagtagccca tgcccaggtc cagccggtac agcatccaga ttccgctgcc
     3241 ggccacgaac acgcagcaca cggcgcaaat gcttcccagc agggggtagt ccatgtcgtt
     3301 gatgctgccc ttgggccagc cctctggagg tctagagtcg tagaaaggca tggtggctag
     3361 cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact
     3421 gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg
     3481 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     3541 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3601 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3661 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3721 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3781 gaactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3841 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3901 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     3961 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4021 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4081 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     4141 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     4201 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     4261 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     4321 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     4381 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     4441 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     4501 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     4561 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     4621 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4681 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4741 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     4801 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     4861 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4921 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4981 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5041 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5101 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5161 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     5221 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5281 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5341 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5401 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5461 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5521 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5581 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5641 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5701 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5761 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5821 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5881 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5941 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6001 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6061 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6121 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6181 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6241 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6301 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6361 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6421 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6481 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6541 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6601 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6661 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6721 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6781 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6841 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6901 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6961 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7021 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7081 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7141 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7201 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7261 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7321 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
