LOCUS       sequence-362032-        7281 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 7281)
  AUTHORS   Kishi KE, Kim YS, Fukuda M, Inoue M, Kusakizako T, Wang PY, 
            Ramakrishnan C, Byrne EFX, Thadhani E, Paggi JM, Matsui TE, 
            Yamashita K, Nagata T, Konno M, Quirin S, Lo M, Benster T, Uemura T,
            Liu K, Shibata M, Nomura N, Iwata S, Nureki O, Dror RO, Inoue K, 
            Deisseroth K, Kato HE
  TITLE     Structural basis for channel conduction in the pump-like 
            channelrhodopsin ChRmine.
  JOURNAL   Cell. 2022 Feb 17;185(4):672-689.e23. doi: 
            10.1016/j.cell.2022.01.007. Epub 2022 Feb 2.
  PUBMED    35114111
REFERENCE   2  (bases 1 to 7281)
  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.2022.01.007"; journalName: "Cell"; date: 
            "2022-02-17- 17"; volume: "185"; issue: "4"; pages: "672-689.e23"
COMMENT     Sequence Label: pAAV-CaMKIIa-ChRmine-oScarlet-KV 2.1-WPRE
FEATURES             Location/Qualifiers
     source          1..7281
                     /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"
     regulatory      1470..1479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2472..3164
                     /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(2602..2620)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2870..2889
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3408..3996
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3461..3481)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3879..3890)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4028..4504
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4227..4246)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4544..4684
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4544..4673
                     /label=AAV2 ITR
     rep_origin      4759..5214
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4846..4865)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5056..5077
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5231..5250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5350..5372
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5410..5428)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5496..5600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5601..5669,5670..6461)
                     /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(5819..5838)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6632..7220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7121..7140
                     /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 cggatccccg ggtaccggtg ccaccatggc acacgcacca ggcaccgacc
     1501 agatgttcta cgtgggcaca atggacggct ggtatctgga taccaagctg aactccgtgg
     1561 ccatcggcgc ccactggtct tgctttatcg tgctgacaat caccacattc tacctgggct
     1621 atgagtcctg gacatctcgg ggccctagca agagaacctc cttttacgcc ggctatcagg
     1681 aggagcagaa cctggccctg tttgtgaatt tctttgccat gctgagctac ttcggcaaga
     1741 tcgtggccga cacactgggc cacaacttcg gcgatgtggg ccccttcatc atcggcttcg
     1801 gcaattacag gtatgccgac tacatgctga cctgcccaat gctggtgtac gatctgctgt
     1861 atcagctgag ggccccctat cgcgtgtctt gtagcgccat catctttgcc atcctgatgt
     1921 ctggcgtgct ggccgagttc tacgcagagg gcgaccctag gctgaggaat ggcgcctacg
     1981 cctggtatgg ctttggctgt ttctggttta tcttcgccta ctctatcgtg atgagcatcg
     2041 tggccaagca gtatagccgg ctggcccagc tggcccagga tacaggcgcc gagcacagcc
     2101 tgcacgtgct gaagttcgcc gtgtttacct tctccatgct gtggattctg tttcccctgg
     2161 tgtgggccat ctgccctaga ggcttcggct ggatcgacga taactggaca gaggtggccc
     2221 actgcgtgtg cgacatcgtg gccaagtcct gttacggctt tgccctggcc cggttcagaa
     2281 agacctatga tgaggagctg tttcggctgc tggagcagct gggacacgac gaggatgagt
     2341 tccagaagct ggagctggat atgaggctga gcagcaatgg cgagcgcctg cggagactgt
     2401 ctgcggccgc caagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg
     2461 acatcaacgt ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     2521 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2581 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     2641 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
     2701 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gaccgcgtga
     2761 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     2821 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     2881 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     2941 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cgggcgctac ctggcggact
     3001 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtggacc
     3061 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     3121 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagcagtcc cagcccatcc
     3181 tcaacaccaa ggagatggcc ccgcagagca agcctccaga ggagctggag atgagcagca
     3241 tgcccagccc cgtggcccct ctgcccgcac gcacggaggg cgtcatcgac atgcggagca
     3301 tgtccagcat tgacagcttc atcagctgtg ccacggactt ccctgaagcc accagattct
     3361 tctgctacga gaacgaggtg taagaattcg atatcaagct tatcgataat caacctctgg
     3421 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3481 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3541 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3601 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggaattg
     3661 ccaccacctg tcagctcctt tccggaactt tcgctttccc cctccctatt gccacggcgg
     3721 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3781 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     3841 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3901 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3961 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga
     4021 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     4081 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     4141 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     4201 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     4261 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     4321 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     4381 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     4441 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     4501 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     4561 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4621 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4681 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4741 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4801 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4861 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4921 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4981 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5041 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     5101 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     5161 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5221 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5281 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5341 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5401 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5461 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5521 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5581 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5641 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5701 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5761 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5821 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5881 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5941 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6001 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6061 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6121 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6181 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6241 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6301 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6361 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6421 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6481 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6541 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6601 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6661 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6721 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6781 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6841 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6901 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6961 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7021 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7081 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7141 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7201 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7261 gctggccttt tgctcacatg t
//
