LOCUS       sequence-406383-        6279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High efficient, K+ selective channelrhodopsin for the inhibition of 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6279)
  AUTHORS   Vierock J, Peter E, Grimm C, Rozenberg A, Chen IW, Tillert L, Castro
            Scalise AG, Casini M, Augustin S, Tanese D, Forget BC, Peyronnet R, 
            Schneider-Warme F, Emiliani V, Beja O, Hegemann P
  TITLE     WiChR, a highly potassium selective channelrhodopsin for low-light 
            one- and two-photon inhibition of excitable cells.
  JOURNAL   Sci Adv. 2022 Nov 16:eadd7729.
  PUBMED    36383037
REFERENCE   2  (bases 1 to 6279)
  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: "Sci Adv. 
            2022 Nov 16:eadd7729."
COMMENT     Sequence Label: pAAV_hSyn_WiChR_TS_mScarlet_ER
FEATURES             Location/Qualifiers
     source          1..6279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..465
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1502..2197
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1635..1653)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1903..1922
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2249..2837
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2302..2322)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2720..2731)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2869..3345
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3068..3087)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3385..3525
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3385..3514
                     /label=AAV2 ITR
     rep_origin      3600..4055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3687..3706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3897..3918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4072..4091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4191..4213
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4251..4269)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4337..4441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4442..4510,4511..5302)
                     /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(4660..4679)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5473..6061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5962..5981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6123..6252
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt
       61 gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt
      121 gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg
      181 ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc
      241 ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc
      301 caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag
      361 ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag
      421 tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgag aaggtaccgg
      481 atccgccacc atgagcgcct gccccacccc ccccaccgtg gtggccccca ccgaccagag
      541 ccccaacccc aacggcggca ccaccagctg cgcccaggcc gcccccagcc cccagatgag
      601 catgaactac accctgctgg ccaccatcga cggcttcagc gccgtgatcc tgctgggcag
      661 cggcgtggtg ttctggtact acagctggcg cggcaccggc ttcggcgccc acccccaccc
      721 caagcgccgc cccctggagc tgaacaccct gagcgccgcc atcgccttcc tgaccggcct
      781 gatgtacgtg gccaagacca gcgacttcgc ccgcgacagc accagcgagt tcagcctgaa
      841 ccgctactgg tacagcgact acatcgtgac ctgccccctg atcgtgatgg acctgtgctt
      901 caccgtggac gtgccctaca agctggcctt caccggcagc accgccgtgc tgctggtgat
      961 cgccctggcc agcttcatcg tgcgcagccc cgagaagtac atctacttcg ccatcggctg
     1021 caccctgttc agcatcttct tctacttctt cttcaacgag gtgcgcaagc gcatccgcta
     1081 cgtgcccgac tgcgccaaga aggacctgga gcgcgccatg ttcatcttct tcggcttctg
     1141 gcccatgttc cccgtgctgt ggctgctggg cttccacacc accggcgtga tcagcgagga
     1201 ggccttcttc atcatccacg ccttcctgga catcacctgc aagaccatct tcggcgtgtt
     1261 catgctgcgc tgccgcctgc acatcgagga ctacgcctgg gacgagctga agcgccagcg
     1321 ccaggagatc gacgaggaga accagaagac cctgaccgcc gccggcgccg acggcgccac
     1381 cgtgatgatg cccgtggagg aggacgaccc ctacgccagc ggacgtacgg cgaagagcag
     1441 gatcaccagc gagggcgagt acatccccct ggaccagatc gacatcaacg tggcggtacc
     1501 catggtgagc aagggcgagg cagtgatcaa ggagttcatg cggttcaagg tgcacatgga
     1561 gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     1621 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tctcctggga
     1681 catcctgtcc cctcagttca tgtacggctc cagggccttc accaagcacc ccgccgacat
     1741 ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1801 cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg gcaccctgat
     1861 ctacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa
     1921 gacaatgggc tgggaagcgt ccaccgagcg gttgtatccc gaggacggcg tgctgaaggg
     1981 cgacattaag atggccctgc gcctgaagga cggcggccgc tacctggcgg acttcaagac
     2041 cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg accgcaagtt
     2101 ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac gctccgaggg
     2161 ccgccactcc accggcggca tggacgagct gtacaagttc tgctacgaga acgaggtgta
     2221 atgagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     2281 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2341 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2401 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2461 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2521 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2581 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2641 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     2701 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2761 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2821 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc
     2881 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     2941 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     3001 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     3061 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     3121 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     3181 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     3241 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     3301 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     3361 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3421 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3481 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3541 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3661 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3781 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     3841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3901 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     3961 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4021 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4081 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4141 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4201 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4261 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4321 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4381 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4441 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4501 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4561 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4621 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4681 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4741 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4801 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4861 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4921 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4981 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5041 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5101 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5161 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5221 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5281 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5341 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5401 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5461 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5521 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5581 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5641 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5701 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5761 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5821 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5881 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5941 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6001 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6061 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6121 gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct
     6181 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     6241 ctaggggttc ctgcggccgc acgcgtgtgt ctagactgc
//
