LOCUS       sequence-390286-        7339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Highly K+ selective channelrhodopsin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7339)
  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 7339)
  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: pCDNA3.1_B1ChR2_TS_mScarlet_ER
FEATURES             Location/Qualifiers
     source          1..7339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        187..566
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        567..770
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     721..741
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     815..834
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        815..833
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2121..2816
                     /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(2254..2272)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2522..2541
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2882..2899)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2888..3112
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3158..3586
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3245..3264)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3455..3476
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3595..3615)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3600..3929
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3780..3915
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3842..3861
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3996..4790
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4050..4069)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4660..4679
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4964..5085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5001..5020)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5055..5074
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5134..5150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5134..5150)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5147..5169)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5158..5174
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5182..5188,5189..5206,5207..5212))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5227..5248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5365..5382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5536..6124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5616..5635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6295..7086,7087..7155))
                     /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     6918..6937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7156..7260)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(7335..7339,1..15))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
       61 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      121 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      181 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      241 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      301 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      361 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      421 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      481 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      541 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      601 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      661 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      721 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      781 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      841 aagctggcta gcgtttagga tccgccacca tgagcgagtg ctgtgaactg tgcgtgtgcc
      901 agaaagagcc tggtacattt ggcgccctga tcgccgtgaa caccatcaca gccattatcc
      961 tggtggccgc tggagcctac atggcctgga agaccgccgc tggcctgggc tggaatacca
     1021 gaccccacgg ccccgagggc cctccagagg aaaactggct gtctcctgga atcagcatcc
     1081 tgtgtggcgt catgtacgcc ttcaaggcca ttgactgggc ctcttataat gataccggag
     1141 agagcaccgc ttttagcctg aaccaggtgt ggtacagcga ctacctgatc acctgtcctt
     1201 tgctggtgct ggacttctgc atcaccgtga acctgcggta caagctggtc ttttctagca
     1261 gcatcgcatg cctgctggcc atcgccgtgt ctacattcat cgtggacgcc ccttacagat
     1321 actacatgta cggcatcggc ctggctggct tcatctgcgc cggctacgcc ctgtggaacg
     1381 agatcaacgc ccaaagagaa aagatccctg actctgcctg gtggtatctg agcgccggaa
     1441 gactgatctt cttcgccgga tggcctttct ttccactgct gtggaccctg agcttccata
     1501 caagcggagt gatcaatgag gagtggtact tcatcctgca cgccatcctg gatatcctgt
     1561 gcaaggctgt tttcggattc ttcatgctcg gctttagact ggaactggag gagctggatt
     1621 tcaaagccat cgaggctgaa caagccaagc tggaaggcga caaggaggcc caggctctta
     1681 aaaacgacaa ggacaccggc gaagtgctga gacactacaa ccgggccagc agcggcttct
     1741 tcggccaggg catccccgac gaccagggct ccgtgtccgg cggcagcgtg ctgatgtcta
     1801 gagcccggcg gcagcacatg ctgtaccggc gcgaagccag ctacctgagc atggacccca
     1861 tggccgagaa gatccgggag ctggagatgc tgaagaagaa gatccagaag gaagtggaca
     1921 gcagaagcaa gaaaatgcag agagagatga cagctagatt cgccatccag gacgactccg
     1981 atgatgaaga tggaggcaga ggcgctggct ccgccaggag aagaaagaac gccagacggg
     2041 gccgtacggc gaagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg
     2101 acatcaacgt ggcggtaccc atggtgagca agggcgaggc agtgatcaag gagttcatgc
     2161 ggttcaaggt gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     2221 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     2281 ccctgccctt ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca
     2341 ccaagcaccc cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt
     2401 gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc
     2461 tggaggacgg caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg
     2521 gccccgtaat gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtatcccg
     2581 aggacggcgt gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct
     2641 acctggcgga cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct
     2701 acaacgtcga ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac
     2761 agtacgaacg ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagttct
     2821 gctacgagaa cgaggtgtaa tgagaattcg atatcaagct taagtttaaa ccgctgatca
     2881 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2941 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3001 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3061 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3121 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3181 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3241 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3301 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3361 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3421 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3481 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3541 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3601 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3661 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3721 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3781 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3841 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3901 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3961 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4021 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4081 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4141 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4201 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4261 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4321 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4381 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4441 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4501 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4561 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4621 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4681 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4741 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4801 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4861 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4921 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4981 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5041 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5101 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5161 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5221 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5281 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5341 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5401 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5461 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5521 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5581 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5641 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5701 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5761 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5821 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5881 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5941 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6001 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6061 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6121 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6181 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6241 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6301 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6361 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6421 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6481 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6541 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6601 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6661 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6721 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6781 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6841 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6901 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6961 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7021 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7081 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7141 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7201 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7261 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7321 tggtgcactc tcagtacaa
//
