LOCUS       sequence-361690-        7538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7538)
  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 7538)
  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 ChRmine-oScarlet-KV 2.1-WPRE
FEATURES             Location/Qualifiers
     source          1..7538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     protein_bind    complement(1476..1509)
                     /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(1560..1593)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1824..2516)
                     /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(2099..2118)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2368..2386
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3509..3518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3523..3556
                     /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    3607..3640
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3665..4253
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3718..3738)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4136..4147)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4285..4761
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4484..4503)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4801..4941
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4801..4930
                     /label=AAV2 ITR
     rep_origin      5016..5471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5103..5122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5313..5334
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5488..5507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5607..5629
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5667..5685)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5753..5857
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5858..5926,5927..6718)
                     /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(6076..6095)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6889..7477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7378..7397
                     /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatcctct agagtcgact ccggaataac ttcgtatagg atactttata
     1501 cgaagttatg cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa
     1561 taacttcgta tagcatacat tatacgaagt tatggcgcgc cctattacac ctcgttctcg
     1621 tagcagaaga atctggtggc ttcagggaag tccgtggcac agctgatgaa gctgtcaatg
     1681 ctggacatgc tccgcatgtc gatgacgccc tccgtgcgtg cgggcagagg ggccacgggg
     1741 ctgggcatgc tgctcatctc cagctcctct ggaggcttgc tctgcggggc catctccttg
     1801 gtgttgagga tgggctggga ctgcttgtac agctcgtcca tgccgccggt ggagtggcgg
     1861 ccctcggagc gttcgtactg ttccaccacg gtgtagtcct cgttgtggga ggtgatgtcc
     1921 aacttgcggt ccacgttgta ggcgccgggc atctgcacgg gcttcttggc cttgtaggtg
     1981 gtcttgaagt ccgccaggta gcgcccgccg tccttcaggc gcagggccat cttaatgtcg
     2041 cccttcagca cgccgtcctc ggggtacaac cgctcggtgg acgcttccca gcccattgtc
     2101 ttcttctgca ttacggggcc gtcaggaggg aagttggtgc cgcggagctt caccttgtag
     2161 atcagggtgc cgtcctccag ggaggtgtcc tgggtcacgg tcacggcgcc gccgtcctcg
     2221 aagttcatca cgcggtccca cttgaagccc tcggggaagg actgcttata gtagtcgggg
     2281 atgtcggcgg ggtgcttggt gaaggccctg gagccgtaca tgaactgagg ggacaggatg
     2341 tcccaggaga agggcagggg gccacccttg gtcaccttca gcttggcggt ctgggtgccc
     2401 tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt catggagccc
     2461 tccatgtgca ccttgaaccg catgaactcc ttgatcactg cctcgccctt gctcaccacg
     2521 ttgatgtcga tctggtccag ggggatgtac tcgccctcgc tggtgatcct gctcttggcg
     2581 gccgcagaca gtctccgcag gcgctcgcca ttgctgctca gcctcatatc cagctccagc
     2641 ttctggaact catcctcgtc gtgtcccagc tgctccagca gccgaaacag ctcctcatca
     2701 taggtctttc tgaaccgggc cagggcaaag ccgtaacagg acttggccac gatgtcgcac
     2761 acgcagtggg ccacctctgt ccagttatcg tcgatccagc cgaagcctct agggcagatg
     2821 gcccacacca ggggaaacag aatccacagc atggagaagg taaacacggc gaacttcagc
     2881 acgtgcaggc tgtgctcggc gcctgtatcc tgggccagct gggccagccg gctatactgc
     2941 ttggccacga tgctcatcac gatagagtag gcgaagataa accagaaaca gccaaagcca
     3001 taccaggcgt aggcgccatt cctcagccta gggtcgccct ctgcgtagaa ctcggccagc
     3061 acgccagaca tcaggatggc aaagatgatg gcgctacaag acacgcgata gggggccctc
     3121 agctgataca gcagatcgta caccagcatt gggcaggtca gcatgtagtc ggcatacctg
     3181 taattgccga agccgatgat gaaggggccc acatcgccga agttgtggcc cagtgtgtcg
     3241 gccacgatct tgccgaagta gctcagcatg gcaaagaaat tcacaaacag ggccaggttc
     3301 tgctcctcct gatagccggc gtaaaaggag gttctcttgc tagggccccg agatgtccag
     3361 gactcatagc ccaggtagaa tgtggtgatt gtcagcacga taaagcaaga ccagtgggcg
     3421 ccgatggcca cggagttcag cttggtatcc agataccagc cgtccattgt gcccacgtag
     3481 aacatctggt cggtgcctgg tgcgtgtgcc atggtggcta gcataacttc gtataaagta
     3541 tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg
     3601 caaagaataa cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat
     3661 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3721 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3781 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3841 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3901 cactggttgg ggcattgcca ccacctgtca gctcctttcc ggaactttcg ctttccccct
     3961 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4021 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     4081 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4141 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4201 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     4261 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     4321 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     4381 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     4441 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     4501 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     4561 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     4621 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     4681 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     4741 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     4801 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4861 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4921 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4981 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     5041 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     5101 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5161 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5221 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5281 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5341 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     5401 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5461 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5581 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5641 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5701 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5761 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5821 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5881 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5941 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6001 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6061 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6121 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6181 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6241 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6301 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6361 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6421 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6481 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6541 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6601 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6661 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6721 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6781 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6841 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6901 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6961 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7021 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7081 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7141 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7201 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7261 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7321 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7381 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7441 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7501 acggttcctg gccttttgct ggccttttgc tcacatgt
//
