LOCUS       sequence-370189-        6333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CapChR1-mScarlet in mammalian neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  AUTHORS   Fernandez Lahore RG, Pampaloni NP, Peter E, Heim MM, Tillert L, 
            Vierock J, Oppermann J, Walther J, Schmitz D, Owald D, Plested AJR, 
            Rost BR, Hegemann P
  TITLE     Calcium-permeable channelrhodopsins for the photocontrol of calcium 
            signalling.
  JOURNAL   Nat Commun. 2022 Dec 21;13(1):7844. doi: 10.1038/s41467-022-35373-4.
  PUBMED    36543773
REFERENCE   2  (bases 1 to 6333)
  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: "Nat 
            Commun."; date: "2022-12-21"; volume: "13(1):7844. doi"; pages: " 
            10.1038/s41467-022-35373-4"
COMMENT     Sequence Label: pAAV-Syn_CapChR1_mScarlet
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      46..501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(133..152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     343..364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(518..537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     637..659
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(697..715)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        783..887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(888..956,957..1748)
                     /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(1106..1125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1919..2507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2408..2427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2569..2698
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2909..3356
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3369..3378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3375..4301
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFDILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVICIHLSNLTGLSNDYSRRTMGLLVSDIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKECWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             4314..5006
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(4444..4462)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4712..4731
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5034..5622
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5087..5107)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5505..5516)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5642..6118
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5841..5860)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   6164..6304
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6164..6293
                     /label=AAV2 ITR
ORIGIN
        1 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
       61 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      121 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      181 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      241 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
      301 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      361 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
      421 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      481 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
      541 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      601 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
      661 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
      721 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
      781 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      841 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      901 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      961 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1021 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1081 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1141 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1201 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1261 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1321 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1381 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1441 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1501 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1561 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1621 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1681 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1741 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1801 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1861 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1921 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1981 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2041 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2101 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2161 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2221 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2281 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2341 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2401 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2461 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2521 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     2581 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     2641 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     2701 ggccgcacgc gttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat
     2761 ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc
     2821 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcagcaaa tggttaatta
     2881 atctagactg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc
     2941 ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca
     3001 ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg
     3061 cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc
     3121 gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc
     3181 cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg
     3241 cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag
     3301 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga
     3361 gaggatccgc caccatggac tatggcggcg ctttgtctgc cgtcggacgc gaacttttgt
     3421 tcgttactaa tcctgtggtg gtgaacgggt ccgtcctggt ccctgaggat caatgttact
     3481 gtgccggatg gattgaatct cgcggcacga acggcgctca gaccgcgtca aatgtcctgc
     3541 agtggcttgc agcaggattc gacattttgc tgctgatgtt ctatgcctac caaacctgga
     3601 aatctacatg cggctgggag gagatctatg tgtgcgccat tgaaatggtt aaggtgattc
     3661 tcgagttctt ttttgagttt aagaatccct ctatgctcta ccttgccaca ggacaccggg
     3721 tgcagtggct gcgctatgca gagtggctgc tcacttgtcc tgtcatctgt atccacctga
     3781 gcaacctcac cggcctgagc aacgactaca gcaggagaac catgggactc cttgtctcag
     3841 acatcgggtg tatcgtgtgg ggggctacca gcgccatggc aaccggctat gttaaagtca
     3901 tcttcttttg tcttggattg tgctatggcg cgaacacatt ttttcacgcc gccaaagcat
     3961 atatcgaggg ttatcatact gtgccaaagg gtcggtgccg ccaggtcgtg accggcatgg
     4021 catggctgtt tttcgtgagc tggggtatgt tcccaattct cttcattttg gggcccgaag
     4081 gttttggcgt cctgagcgtc tatggctcca ccgtaggtca cacgattatt gatctgatga
     4141 gtaaagagtg ttgggggttg ttgggacact acctgcgcgt cctgatccac gagcacatat
     4201 tgattcacgg agatatccgc aaaaccacca aactgaacat cggcggaacg gagatcgagg
     4261 tcgagactct cgtcgaagac gaagccgagg ccggagccgt gccagcggcc gccgtgagca
     4321 agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga
     4381 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     4441 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc
     4501 ctcagttcat gtacggctcc agggccttca ccaagcaccc cgccgacatc cccgactact
     4561 ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     4621 gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc tacaaggtga
     4681 agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct
     4741 gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc gacattaaga
     4801 tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc acctacaagg
     4861 ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg gacatcacct
     4921 cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc cgccactcca
     4981 ccggcggcat ggacgagctg tacaagtaag aattccgtac aaagcttatc gataatcaac
     5041 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5101 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5161 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5221 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5281 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5341 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5401 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     5461 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5521 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     5581 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cggctcgaga
     5641 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     5701 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     5761 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     5821 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     5881 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     5941 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     6001 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     6061 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     6121 cgagctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     6181 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     6241 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     6301 caggggcgcc tgatgcggta ttttctcctt acg
//
