LOCUS       sequence-182715-        5635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  blue-shifted artificial anion conducting channelrhodopsin (aACR). 
            Activation max 466 nm; off-kinetics 10 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-Phobos
FEATURES             Location/Qualifiers
     source          1..5635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        118..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1599..2306
                     /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(1741..1759)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2009..2028
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2480..2601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2517..2536)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2571..2590
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2608..3063)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2745..2766)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2957..2976
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3090..3194
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3196..3553
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3196..3387
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3219..3239)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3404..3539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3588..4382
                     /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             3588..4382
                     /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(3642..3661)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4252..4271
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4570..4589)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4614..4661
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4990..5578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5479..5498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgccacca
      661 tggactatgg cggcgctttg tctgccgtcg gactcttcca aacctcatac actcttgaga
      721 acaatggttc tgttatttgc atcccgaaca acggccagtg cttctgcttg gcttggctta
      781 aatccaacgg aacaaatgcc gagaagttgg ctgccaacat tctgcagtgg atttcttttg
      841 cgctttcagc gctctgcctg atgttctacg gctaccagac ctggaagtct acttgcggct
      901 gggagaacat ttacgtggcc acgatccaga tgatcaagtt catcatcgag tatttccata
      961 gctttgacga acctgcggtg atctactcat ccaacggcaa caagacccgg tggcttcgtt
     1021 acgcgagctg gctgctgacc tgccctgtca ttcttatcca cctgagcaac cttacgggtc
     1081 tggcgaacga ctataacaag cgtaccatgg gtctgctggt gtcagatatc ggcggcatcg
     1141 tgtgggccac cacggccgcg ctgtccaagg gatacgtccg tgtcattttc ttcctgatgg
     1201 gcctgtgcta cggcatctac acattcttca acgcagccaa ggtctacatt gaggcgtacc
     1261 acaccgtgcc gaagggccgg tgtcgccagg tggtgactgg catggcttgg ctcttcttcg
     1321 tatcatgggg tatgttcccc atcctgttca tcctcggccc cgagggcttc ggcgtcctga
     1381 gccggtacgg ctccaacgtc ggccacacca tcattgacct gatgtcgaag cagtgctggg
     1441 gtctgctcgg ccactacctg cgcgtgctga tccacagcca tatcctcatc cacggcgaca
     1501 ttcgcaagac caccaaattg aacattggtg gcactgagat tgaggtcgag acgctggtgg
     1561 aggacgaggc cgaggctggc gcggtcgcgg caccggtggt gagcaagggc gaggaggata
     1621 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     1681 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1741 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1801 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     1861 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1921 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1981 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2041 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     2101 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     2161 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     2221 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     2281 gcggcatgga cgagctgtac aagtaagagc tcaagcttcg aattctgcag tcgacggtac
     2341 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     2401 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2461 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2521 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2581 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2641 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2701 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2761 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2821 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2881 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2941 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3001 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3181 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3301 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3361 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3421 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3481 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3541 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3601 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3661 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3721 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3781 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3841 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3901 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3961 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4021 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4081 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4141 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4201 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4261 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4321 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4381 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4441 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4501 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4561 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4621 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4681 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4741 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4801 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4861 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4921 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4981 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5041 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5101 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5161 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5221 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5281 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5341 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5401 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5461 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5521 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5581 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcac
//
