LOCUS       sequence-166563-        5588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Natural anion conducting Channelrhodopsin with cyan absorption. 
            Closing kinetics 140ms. High conductivity. Codon optimized for 
            mammalian expression (human/mouse).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5588)
  AUTHORS   Wietek J, Broser M, Krause BS, Hegemann P
  TITLE     Identification of a Natural Green Light Absorbing Chloride 
            Conducting Channelrhodopsin from Proteomonas sulcata.
  JOURNAL   J Biol Chem. 2016 Feb 19;291(8):4121-7. doi: 
            10.1074/jbc.M115.699637. Epub 2016 Jan 6.
  PUBMED    26740624
REFERENCE   2  (bases 1 to 5588)
  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"; doi: 
            "10.1074/jbc.M115.699637"; journalName: "J Biol Chem"; date: 
            "2016-02-19- 19"; volume: "291"; issue: "8"; pages: "4121-7"
COMMENT     Sequence Label: p-mCherry-C1-GtACR1
FEATURES             Location/Qualifiers
     source          1..5588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        45..348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        349..552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     503..523
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1479..2186
                     /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(1621..1639)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1889..1908
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2360..2481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2397..2416)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2451..2470
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2488..2943)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2625..2646)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2837..2856
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2970..3074
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3076..3433
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3076..3267
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3099..3119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3284..3419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3468..4262
                     /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             3468..4262
                     /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(3522..3541)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4132..4151
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4450..4469)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4494..4541
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4870..5458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5359..5378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
       61 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      121 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      181 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      241 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      301 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      361 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      421 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      481 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      541 ataagcagag ctggtttagt gaaccgtcag atccgctagc caccatgagc agcattactt
      601 gtgacccagc catctatgga gagtggagca gagagaacca gttttgcgtc gaaaagagcc
      661 tgattaccct ggacggcatc aagtacgtgc agctggtcat ggccgtggtg agcgcctgcc
      721 aggtgttctt tatggtcact agggcaccca aagtgccttg ggaggccatc tacctgccaa
      781 ccacagaaat gatcacctat tcactggctt tcacaggcaa cggatacatt cgagtggcaa
      841 atgggaagta tctgccttgg gctagaatgg caagttggct gtgcacatgt ccaatcatgc
      901 tgggcctggt gagcaacatg gccctggtga agtacaaatc cattccactg aatcccatga
      961 tgattgccgc cagcagcatc tgcacagtgt tcggaatcac tgcctccgtg gtcctggacc
     1021 ccctgcacgt gtggctgtac tgtttcatct ctagtatttt ctttatcttt gagatggtgg
     1081 tcgcattcgc catttttgcc atcactattc atgattttca gaccatcgga agtcccatgt
     1141 cactgaaggt ggtcgaaagg ctgaaactga tgagaatcgt gttctacgtg agctggatgg
     1201 cctatcctat cctgtggagt ttttcaagca caggggcttg cattatgtct gagaacactt
     1261 cctctgtgct gtacctgctg ggcgacgcac tgtgcaaaaa tacctatgga atcctgctgt
     1321 gggctactac ctggggactg ctgaacggca agtgggaccg agattacgtg aaaggacgca
     1381 atgtcgatgg gacactgatg cccgaatatg aacaggatct ggagaaaggc aatactgaac
     1441 gatatgagga tgctcgggct ggggaaaccg gaccggtggt gagcaagggc gaggaggata
     1501 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     1561 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1621 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1681 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     1741 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1801 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1861 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1921 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     1981 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     2041 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     2101 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     2161 gcggcatgga cgagctgtac aagtaagagc tcaagcttcg aattctgcag tcgacggtac
     2221 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     2281 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2341 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2401 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2461 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2521 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2581 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2641 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2701 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2761 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2821 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2881 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2941 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3001 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3061 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3121 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3181 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3241 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3301 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3361 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3421 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3481 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3541 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3601 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3661 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3721 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3781 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3841 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3901 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3961 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4021 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4081 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4141 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4201 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4261 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4321 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4381 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4441 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4501 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4561 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4621 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4681 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4741 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4801 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4981 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5521 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     5581 aatagtaa
//
