LOCUS       sequence-170779-        5576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Natural anion conducting Channelrhodopsin with red shifted 
            absorption. Closing kinetics 80ms. 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-PsACR1
FEATURES             Location/Qualifiers
     source          1..5576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..406
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        49..240
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(72..92)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      257..392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             441..1235
                     /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             441..1235
                     /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(495..514)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1105..1124
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1423..1442)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1467..1514
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1843..2431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2332..2351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2606..2909
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2910..3113
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3064..3084
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4028..4735
                     /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(4170..4188)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4438..4457
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4909..5030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4946..4965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5000..5019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5037..5492)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5174..5195)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5386..5405
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5519..5576,1..47)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
       61 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
      121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
      181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
      241 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
      301 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
      361 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
      421 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
      481 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
      541 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
      601 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
      661 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
      721 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
      781 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
      841 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
      901 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
      961 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     1021 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     1081 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     1141 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     1201 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     1261 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     1321 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     1381 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     1441 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     1501 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     1561 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     1621 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     1681 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1741 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1801 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1981 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2461 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2521 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     2581 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2641 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2701 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2761 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2821 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2881 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2941 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     3001 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     3061 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     3121 tgaaccgtca gatccgctag ccaccatgac aactatctcc gaggtctgtg gggtctgggc
     3181 actggacaat cccgaatgta tcgaagtctc aggaacaaac gacaatgtga aaatggccca
     3241 gctgtgcttc tgtatggtgt gcgtctgtca gatcctgttt atggccagcc agtacccaaa
     3301 agtgggatgg gaggcaatct acctgccctc ctgcgaatgt ttcctgtacg ggctggccag
     3361 ctccggaaac ggcttcatcc agctgtatga cggccgactg attccttggg ctagatacgc
     3421 cgcttggatc tgcacttgtc catctattct gctgcagatc aataccattc acaagtgtaa
     3481 aatcagtcat ttcaacctga ataccttcat cgtgcaggca gatctgatca tgaacattat
     3541 gggggtcact ggcgcactga ccacaaatat cgccttcaag tggatctact ttgccatcgg
     3601 ctgcattctg ttcatcttta ttgtgctggt ggtctacgac atcatgacct ctgcagccaa
     3661 ggagtggaag gctaaaggag attccaaagg gaacctggtg tctacacggc tgatcctgct
     3721 gcgctggatc ttcattgtga gttggtgcgt ctatcccctg ctgtggattc tgtcacctca
     3781 ggccacttgt gccgtgagcg aagacgtgat cagcgtcgct cacttcattt gcgatgcttt
     3841 tgcaaagaac atgttcggct ttatcatgtg gaggacactg tggagagacc tggatggaca
     3901 ctgggacatt agcaggcatt acccccagtc tagttatgca aaggatggca aagaggaaga
     3961 acagatgact gctatgagcc agactgatga cactgaaaaa ccccattcca gccagggcgg
     4021 accggtggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     4081 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     4141 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     4201 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     4261 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     4321 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     4381 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     4441 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     4501 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     4561 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     4621 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     4681 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagagct
     4741 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa
     4801 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4861 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4921 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4981 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     5041 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5101 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5161 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5221 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     5281 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5341 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5401 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5461 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     5521 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatga
//
