LOCUS       sequence-170777-        5576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Natural anion conducting Channelrhodopsin with blue absorption. 
            Closing kinetics 60ms. 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 5576)
  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 5576)
  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-GtACR2
FEATURES             Location/Qualifiers
     source          1..5576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..453
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        96..287
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(119..139)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      304..439
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             488..1282
                     /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             488..1282
                     /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(542..561)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1152..1171
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1470..1489)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1514..1561
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1890..2478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2379..2398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2653..2956
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2957..3160
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3111..3131
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3187..3196
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4075..4782
                     /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(4217..4235)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4485..4504
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4956..5077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4993..5012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5047..5066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5084..5539)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5221..5242)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5433..5452
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5566..5576,1..94)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
       61 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
      121 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      181 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      241 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      301 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      361 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
      421 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
      481 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
      541 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
      601 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
      661 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
      721 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
      781 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
      841 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
      901 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
      961 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     1021 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     1081 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     1141 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     1201 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     1261 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     1321 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     1381 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     1441 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     1501 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     1561 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     1621 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     1681 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     1741 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1801 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1861 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1921 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1981 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2041 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2101 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2161 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2221 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2281 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2341 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2401 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2461 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2521 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2581 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     2641 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     2701 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2761 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2821 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2881 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2941 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3001 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     3061 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     3121 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     3181 ccgctagcca ccatggcatc acaggtcgtc tatggggaat gggcatcaac tcacaccgag
     3241 tgctacaata tgtccagaat cgactccacc ttcgtctcac tgctgcagct ggtgtgggcc
     3301 gtggtctcag gctgccagac aatcttcatg attagcaggg ctcccaaggt cccttgggag
     3361 tccgtgtacc tgccctttgt cgaatctatc acttatgcac tggccagtac tggcaacgga
     3421 accctgcaga tgagggacgg cagattcttt ccttggagcc gaatggcctc ctggctgtgc
     3481 acctgtccaa tcatgctggg acagatttct aacatggctc tggtgaagta caaaagtatc
     3541 ccactgaatc ccattgcaca ggccgcttcc atcattagag tggtcatggg aatcaccgct
     3601 acaatttctc ccgcagagta tatgaagtgg ctgttctttt tctttggggc cacctgcctg
     3661 gtgttcgagt acagtgtggt cttcacaatc tttcaagtgg ggctgtatgg ctttgaatct
     3721 gtcgggacac ctctggcaca gaaggtggtg gtgaggatca agatgctgcg cctgatcttc
     3781 tttattgctt ggactatgtt cccaatcgtg tggctgattt cacccaccgg agtctgcgtg
     3841 atccacgaga acgtgagcgc cattctgtac ctgctggctg acggcctgtg caaaaataca
     3901 tatggagtga tcctgtggag cactgcctgg ggagtgctgg aagggaagtg ggaccccgca
     3961 tgtctgccag ggcaggagaa acccgaagcc gacgatcctt ttggcctgaa tcacgagaag
     4021 aatgcccctc ctaacgacga agtcaatatc agaatgtttg gaaggggacc ggtggtgagc
     4081 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     4141 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     4201 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     4261 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     4321 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     4381 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     4441 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     4501 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     4561 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     4621 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     4681 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     4741 ggccgccact ccaccggcgg catggacgag ctgtacaagt aagagctcaa gcttcgaatt
     4801 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     4861 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4921 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4981 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5041 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     5101 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5161 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5221 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5281 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5341 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5401 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5461 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5521 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccc
//
