LOCUS       sequence-182770-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  artificial anion conducting channelrhodopsin (aACR). Activation with
            blue light. Codon optimized for mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  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 5630)
  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-iChR2++
FEATURES             Location/Qualifiers
     source          1..5630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        127..430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        431..634
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     585..605
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      661..670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1603..2310
                     /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(1745..1763)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2013..2032
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2484..2605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2521..2540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2575..2594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2612..3067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2749..2770)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2961..2980
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3094..3198
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3200..3557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3200..3391
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3223..3243)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3408..3543
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3592..4386
                     /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             3592..4386
                     /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(3646..3665)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4256..4275
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4574..4593)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4618..4665
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4994..5582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5483..5502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
       61 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      601 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      661 gccaccatgg attatggggg agcactgtca gctgtgggcc tgtttcagac ttcctacacc
      721 ctggagaaca atgggagcgt catctgtatt cctaacaatg gccagtgctt ttgtctggcc
      781 tggctgaagt ctaacggcac caatggcgcc cagacagcca gcaacgtgct gcagtggctg
      841 agcgccggct tttccatcct gctgctgatg ttctacgcct atcagacctg gaagtccaca
      901 tgcggctggg agaacatcta cgtgtgcgcc atccagatgg tgaaagtgat cctggagttc
      961 tttttcagct tcaagaatcc atccatgctg tacctggcaa ccggccaccg ggtgcgctgg
     1021 ctgagatatg ccagctggct gctgacatgc cccgtgatcc tgatccacct gtccaacctg
     1081 accggcctgt ctaatgacta tagcaggaga acaatgggcc tgctggtgtc cgatatcggc
     1141 tgtatcgtgt ggggagccac ctctgccatg gcaacaggct acgtgaaagt gatcttcttc
     1201 tgcctgggcc tgtgctatgg cgccaatacc tttttccacg ccgccaaggc ctacatcgag
     1261 ggctatcaca ccgtgcccaa gggcaggtgc agacaggtgg tgacaggcat ggcctggctg
     1321 tttttcgtgt cttggggcat gtttcccatc ctgttcatcc tgggccctga gggctttggc
     1381 gtgctgtcca ggtacggctc taacgtgggc cacacaatca tcgacctgat gtctaagcag
     1441 tgttggggcc tgctgggaca ctatctgcgg gtgctgatcc acagccacat cctgatccac
     1501 ggcgatatcc gcaagaccac aaagctgaat atcggaggga ccgagattga agtggaaact
     1561 ctggtggaag acgaagccga agcaggagca gtgggaccgg tggtgagcaa gggcgaggag
     1621 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1681 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1741 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1801 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     1861 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     1921 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     1981 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2041 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2101 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2161 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2221 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2281 accggcggca tggacgagct gtacaagtaa gagctcaagc ttcgaattct gcagtcgacg
     2341 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     2401 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2461 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2521 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2581 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     2641 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2701 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2761 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2821 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2881 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2941 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3001 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3061 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3121 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3181 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3301 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3361 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3421 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3481 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3541 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3601 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3661 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3721 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3781 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3841 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3901 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3961 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4021 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4081 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4141 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4201 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4261 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4321 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4381 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4441 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4501 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4561 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4621 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4681 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4741 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4801 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4861 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4921 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4981 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5041 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5101 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5161 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5221 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5281 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5341 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5401 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5461 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5521 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5581 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
//
