LOCUS       sequence-170772-        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"
     promoter        77..181
                     /gene="bla"
                     /label=AmpR promoter
     promoter        183..540
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        183..374
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(206..226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      391..526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             575..1369
                     /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             575..1369
                     /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(629..648)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1239..1258
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1557..1576)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1601..1648
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1977..2565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2466..2485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2740..3043
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3044..3247
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3198..3218
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4174..4881
                     /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(4316..4334)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4584..4603
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5055..5176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5092..5111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5146..5165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(5183..5588,1..50))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5320..5341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5532..5551
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
       61 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      121 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      181 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
      241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
      301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
      361 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
      421 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
      481 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
      541 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
      601 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
      661 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
      721 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
      781 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
      841 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
      901 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
      961 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     1021 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     1081 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     1141 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     1201 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     1261 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     1321 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     1381 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     1441 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     1501 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     1561 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     1621 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     1681 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     1741 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     1801 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     1861 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1921 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1981 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2041 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2101 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2161 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2221 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2281 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2341 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2401 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2461 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2521 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2581 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2641 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     2701 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     2761 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     2821 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     2881 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     2941 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     3001 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     3061 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     3121 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     3181 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     3241 cagagctggt ttagtgaacc gtcagatccg ctagccacca tgagcagcat tacttgtgac
     3301 ccagccatct atggagagtg gagcagagag aaccagtttt gcgtcgaaaa gagcctgatt
     3361 accctggacg gcatcaagta cgtgcagctg gtcatggccg tggtgagcgc ctgccaggtg
     3421 ttctttatgg tcactagggc acccaaagtg ccttgggagg ccatctacct gccaaccaca
     3481 gaaatgatca cctattcact ggctttcaca ggcaacggat acattcgagt ggcaaatggg
     3541 aagtatctgc cttgggctag aatggcaagt tggctgtgca catgtccaat catgctgggc
     3601 ctggtgagca acatggccct ggtgaagtac aaatccattc cactgaatcc catgatgatt
     3661 gccgccagca gcatctgcac agtgttcgga atcactgcct ccgtggtcct ggaccccctg
     3721 cacgtgtggc tgtactgttt catctctagt attttcttta tctttgagat ggtggtcgca
     3781 ttcgccattt ttgccatcac tattcatgat tttcagacca tcggaagtcc catgtcactg
     3841 aaggtggtcg aaaggctgaa actgatgaga atcgtgttct acgtgagctg gatggcctat
     3901 cctatcctgt ggagtttttc aagcacaggg gcttgcatta tgtctgagaa cacttcctct
     3961 gtgctgtacc tgctgggcga cgcactgtgc aaaaatacct atggaatcct gctgtgggct
     4021 actacctggg gactgctgaa cggcaagtgg gaccgagatt acgtgaaagg acgcaatgtc
     4081 gatgggacac tgatgcccga atatgaacag gatctggaga aaggcaatac tgaacgatat
     4141 gaggatgctc gggctgggga aaccggaccg gtggtgagca agggcgagga ggataacatg
     4201 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     4261 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     4321 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     4381 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     4441 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     4501 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     4561 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     4621 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     4681 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     4741 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     4801 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     4861 atggacgagc tgtacaagta agagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
     4921 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     4981 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     5041 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5101 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5161 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     5221 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5281 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5341 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5401 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5461 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5521 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5581 gtcacgct
//
