LOCUS       sequence-370192-        6297 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses CapChR2-eYFP in mammalian neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  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 6297)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_CapChR2_eYFP
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..717)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(43..64)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(304..323)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     651..672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    742..1330
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(795..815)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1213..1224)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1350..1826
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1549..1568)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   1872..2012
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1872..2001
                     /label=AAV2 ITR
     rep_origin      2087..2542
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2174..2193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2384..2405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2559..2578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2678..2700
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2738..2756)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2824..2928
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2929..2997,2998..3789)
                     /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(3147..3166)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3960..4548
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4449..4468
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4610..4739
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        4950..5397
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             5416..6279
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFDIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVICIHLSNLTGLKDDYSKRTMRLLVSDVGCIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKECWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
ORIGIN
        1 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
       61 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      121 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      181 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
      241 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      301 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      361 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      421 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
      481 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
      541 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      601 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      661 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagaa
      721 ttccgtacaa agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
      781 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
      841 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
      901 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
      961 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     1021 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     1081 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     1141 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     1201 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     1261 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     1321 gcctccccgc atcgataccg gctcgagacg ggtggcatcc ctgtgacccc tccccagtgc
     1381 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     1441 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     1501 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     1561 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     1621 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     1681 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     1741 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     1801 gaaccactgc tcccttccct gtccttctcg agctgatttt gtaggtaacc acgtgcggac
     1861 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     1921 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     1981 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     2041 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     2101 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2161 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2221 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2281 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2341 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2401 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     2461 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2521 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     2581 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2641 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2701 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     2761 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2821 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2881 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2941 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3001 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3061 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3121 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3181 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3241 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3301 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3361 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3421 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3481 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3541 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3601 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3661 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3721 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3781 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3841 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3901 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3961 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4021 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4081 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4141 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4201 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4261 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4321 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4381 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4441 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4501 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4561 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag
     4621 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
     4681 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
     4741 cggccgcacg cgttgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa
     4801 tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag
     4861 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcagcaa atggttaatt
     4921 aatctagact gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg
     4981 cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc
     5041 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc
     5101 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg
     5161 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact
     5221 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac
     5281 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca
     5341 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg
     5401 agaggatccg ccaccatgct gggaaacggc agcgccattg tgcctatcga ccagtgcttt
     5461 tgcctggctt ggaccgacag cctgggaagc gatacagagc agctggtggc caacatcctc
     5521 cagtggttcg ccttcggctt cgacatcctg atcctgatgt tctacgccta ccagacttgg
     5581 agagccactt gcggttggga ggaggtctac gtctgttgcg tcgagctgac caaggtcatc
     5641 atcgagttct tccacgagtt cgacgacccc agcatgctgt acctggctaa cggacaccga
     5701 gtccagtggc tgagatacgc agagtggctg ctgacttgtc ccgtcatctg catccacctg
     5761 agcaacctga ccggcctgaa ggacgactac agcaagcgga ccatgaggct gctggtgtca
     5821 gacgtgggat gcatcgtgtg gggagctaca agcgccatga gcacaggcta cgtcaaggtc
     5881 atcttcttcg tgctgggttg catctacggc gccaacacct tcttccacgc cgccaaggtg
     5941 tatatcgaga gctaccacgt ggtgccaaag ggcagaccta gaaccgtcgt gcggatcatg
     6001 gcttggctgt tcttcctgtc ttggggcatg ttccccgtgc tgttcgtcgt gggaccagaa
     6061 ggattcgacg ccatcagcgt gtacggctct accattggcc acaccatcat cgacctcatg
     6121 agcaaggagt gttggggcct gctgggacac tatctgagag tgctgatcca ccagcacatc
     6181 atcatctacg gcgacatccg gaagaagacc aagatcaacg tggccggcga ggagatggaa
     6241 gtggagacca tggtggacca ggaggacgag gagacagtgg cggcaccggt agtagca
//
