LOCUS       sequence-182759-        5635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  slow cycling (open for minutes) step-function artificial anion 
            conducting channelrhodopsin (aACR). High light sensitivity. 
            Activation with cyan light, inactivation max with 590 nm 
            (accelarates closure to ms). Codon optimized for mammalian 
            expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5635)
  AUTHORS   Wietek J, Rodriguez-Rozada S, Tutas J, Tenedini F, Grimm C, Oertner 
            TG, Soba P, Hegemann P, Wiegert JS
  TITLE     Anion-conducting channelrhodopsins with tuned spectra and modified 
            kinetics engineered for optogenetic manipulation of behavior.
  JOURNAL   Sci Rep. 2017 Nov 2;7(1):14957. doi: 10.1038/s41598-017-14330-y.
  PUBMED    29097684
REFERENCE   2  (bases 1 to 5635)
  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 Rep."; 
            date: "2017-11-2"; volume: "7(1):14957. doi"; pages: " 
            10.1038/s41598-017-14330-y"
COMMENT     Sequence Label: p-mCherry-C1-iC++_CA
FEATURES             Location/Qualifiers
     source          1..5635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        91..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        395..598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     549..569
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      627..636
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1572..2279
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1714..1732)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1982..2001
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2453..2574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2490..2509)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2544..2563
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2581..3036)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2718..2739)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2930..2949
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3063..3167
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3169..3526
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3169..3360
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3192..3212)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3377..3512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3561..4355
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3561..4355
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3615..3634)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4225..4244
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4543..4562)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4587..4634
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4963..5551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5452..5471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
       61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      421 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      481 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      541 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      601 tttagtgaac cgtcagatcc gctagcgcca ccatggacta tggcggcgct ttgtctgccg
      661 tcggactctt ccaaacctca tacactcttg agaacaatgg ttctgttatt tgcatcccga
      721 acaacggcca gtgcttctgc ttggcttggc ttaaatccaa cggaacaaat gccgagaagt
      781 tggctgccaa cattctgcag tggatttctt ttgcgctttc agcgctctgc ctgatgttct
      841 acggctacca gacctggaag tctacttgcg gctgggagaa catttacgtg gccacgatcc
      901 agatgatcaa gttcatcatc gagtatttcc atagctttga cgaacctgcg gtgatctact
      961 catccaacgg caacaagacc cggtggcttc gttacgcgag ctggctgctg accgcccctg
     1021 tcattcttat ccacctgagc aaccttacgg gtctggcgaa cgactataac aagcgtacca
     1081 tgggtctgct ggtgtcagat atcggcacga tcgtgtgggg caccacggcc gcgctgtcca
     1141 agggatacgt ccgtgtcatt ttcttcctga tgggcctgtg ctacggcatc tacacattct
     1201 tcaacgcagc caaggtctac attgaggcgt accacaccgt gccgaagggc cggtgtcgcc
     1261 aggtggtgac tggcatggct tggctcttct tcgtatcatg gggtatgttc cccatcctgt
     1321 tcatcctcgg ccccgagggc ttcggcgtcc tgagccggta cggctccaac gtcggccaca
     1381 ccatcattga cctgatgtcg aagcagtgct ggggtctgct cggccactac ctgcgcgtgc
     1441 tgatccacag ccatatcctc atccacggcg acattcgcaa gaccaccaaa ttgaacattg
     1501 gtggcactga gattgaggtc gagacgctgg tggaggacga ggccgaggct ggcgcggtcg
     1561 cggcaccggt ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     1621 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     1681 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     1741 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     1801 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     1861 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     1921 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     1981 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     2041 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     2101 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2161 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     2221 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag
     2281 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggatctag
     2341 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2401 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2461 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2521 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     2581 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2641 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2701 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2761 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2821 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2881 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2941 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3001 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3061 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3121 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3181 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3301 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3361 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3421 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3481 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3541 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3601 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3661 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3721 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3781 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3841 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3901 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3961 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4021 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4081 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4141 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4201 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4261 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4321 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4381 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4441 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4501 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4561 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4621 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4681 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4741 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4801 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4861 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4921 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4981 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5041 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5101 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5161 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5221 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5281 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5341 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5401 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5461 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5521 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5581 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgat
//
