LOCUS       sequence-166564-        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"
     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"
     regulatory      579..588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1467..2174
                     /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(1609..1627)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1877..1896
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2348..2469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2385..2404)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2439..2458
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2476..2931)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2613..2634)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2825..2844
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2958..3062
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3064..3421
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3064..3255
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3087..3107)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3272..3407
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3456..4250
                     /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             3456..4250
                     /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(3510..3529)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4120..4139
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4438..4457)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4482..4529
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4858..5446
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5347..5366
                     /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 caccatggca tcacaggtcg
      601 tctatgggga atgggcatca actcacaccg agtgctacaa tatgtccaga atcgactcca
      661 ccttcgtctc actgctgcag ctggtgtggg ccgtggtctc aggctgccag acaatcttca
      721 tgattagcag ggctcccaag gtcccttggg agtccgtgta cctgcccttt gtcgaatcta
      781 tcacttatgc actggccagt actggcaacg gaaccctgca gatgagggac ggcagattct
      841 ttccttggag ccgaatggcc tcctggctgt gcacctgtcc aatcatgctg ggacagattt
      901 ctaacatggc tctggtgaag tacaaaagta tcccactgaa tcccattgca caggccgctt
      961 ccatcattag agtggtcatg ggaatcaccg ctacaatttc tcccgcagag tatatgaagt
     1021 ggctgttctt tttctttggg gccacctgcc tggtgttcga gtacagtgtg gtcttcacaa
     1081 tctttcaagt ggggctgtat ggctttgaat ctgtcgggac acctctggca cagaaggtgg
     1141 tggtgaggat caagatgctg cgcctgatct tctttattgc ttggactatg ttcccaatcg
     1201 tgtggctgat ttcacccacc ggagtctgcg tgatccacga gaacgtgagc gccattctgt
     1261 acctgctggc tgacggcctg tgcaaaaata catatggagt gatcctgtgg agcactgcct
     1321 ggggagtgct ggaagggaag tgggaccccg catgtctgcc agggcaggag aaacccgaag
     1381 ccgacgatcc ttttggcctg aatcacgaga agaatgcccc tcctaacgac gaagtcaata
     1441 tcagaatgtt tggaagggga ccggtggtga gcaagggcga ggaggataac atggccatca
     1501 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     1561 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     1621 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     1681 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1741 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1801 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1861 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1921 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1981 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     2041 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     2101 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     2161 agctgtacaa gtaagagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg
     2221 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2281 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2341 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2401 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2461 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2521 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2581 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2641 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2701 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2761 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2821 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2881 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2941 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3001 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3061 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3241 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3301 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3361 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3421 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3481 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3541 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3601 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3661 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3721 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3781 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3841 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3901 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3961 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4021 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4081 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4141 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4201 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4261 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4321 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4381 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4441 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4501 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4561 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4621 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4681 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4741 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4801 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4861 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4921 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4981 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5041 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5101 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5161 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5221 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5281 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5341 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5401 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5461 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5521 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaa
//
