LOCUS       sequence-170773-        5659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  improved slowChloC artificial anion conducting ChR. Codon optimized 
            for mammalian expression (human/mouse).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5659)
  AUTHORS   Wietek J, Beltramo R, Scanziani M, Hegemann P, Oertner TG, Simon 
            Wiegert J
  TITLE     An improved chloride-conducting channelrhodopsin for light-induced 
            inhibition of neuronal activity in vivo.
  JOURNAL   Sci Rep. 2015 Oct 7;5:14807. doi: 10.1038/srep14807.
  PUBMED    26443033
REFERENCE   2  (bases 1 to 5659)
  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"; journalName: "Sci Rep."; 
            date: "2015-10-7"; volume: "5:14807. doi"; pages: " 
            10.1038/srep14807"
COMMENT     Sequence Label: p-mCherry-N1-iChloC
FEATURES             Location/Qualifiers
     source          1..5659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        56..247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(79..99)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      264..399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             448..1242
                     /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             448..1242
                     /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(502..521)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1112..1131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1430..1449)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1474..1521
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1850..2438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2339..2358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2613..2916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2917..3120
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3071..3091
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3143..3223
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      3224..3233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3230..4156
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEQIYVCAIRMVKVILEFFFSFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSNIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             4169..4876
                     /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(4311..4329)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4579..4598
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4999..5120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5036..5055)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5090..5109
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5127..5582)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5264..5285)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5476..5495
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5609..5659,1..54)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
       61 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
      361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
      421 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
      481 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
      541 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
      601 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
      661 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
      721 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
      781 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
      841 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
      901 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
      961 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     1021 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     1081 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     1141 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     1201 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     1261 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     1321 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     1381 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     1441 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     1501 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1561 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1621 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     1681 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1981 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2461 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2521 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     2581 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2641 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2701 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2761 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2821 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2881 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2941 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     3001 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     3061 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     3121 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc
     3181 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtgccacca tggactatgg
     3241 cggcgctttg tctgccgtcg gacgcgaact tttgttcgtt actaatcctg tggtggtgaa
     3301 cgggtccgtc ctggtccctg aggatcaatg ttactgtgcc ggatggattg aatctcgcgg
     3361 cacgaacggc gctcagaccg cgtcaaatgt cctgcagtgg cttgcagcag gattcagcat
     3421 tttgctgctg atgttctatg cctaccaaac ctggaaatct acatgcggct gggagcagat
     3481 ctatgtgtgc gccattagaa tggttaaggt gattctcgag ttctttttta gctttaagaa
     3541 tccctctatg ctctaccttg ccacaggaca ccgggtgcag tggctgcgct atgcagagtg
     3601 gctgctcact tgtcctgtca tccttatcca cctgagcaac ctcaccggcc tgagcaacga
     3661 ctacagcagg agaaccatgg gactccttgt ctcaaacatc gggtgtatcg tgtggggggc
     3721 taccagcgcc atggcaaccg gctatgttaa agtcatcttc ttttgtcttg gattgtgcta
     3781 tggcgcgaac acattttttc acgccgccaa agcatatatc gagggttatc atactgtgcc
     3841 aaagggtcgg tgccgccagg tcgtgaccgg catggcatgg ctgtttttcg tgagctgggg
     3901 tatgttccca attctcttca ttttggggcc cgaaggtttt ggcgtcctga gcgtctatgg
     3961 ctccaccgta ggtcacacga ttattgatct gatgagtaaa aattgttggg ggttgttggg
     4021 acactacctg cgcgtcctga tccacgagca catattgatt cacggagata tccgcaaaac
     4081 caccaaactg aacatcggcg gaacggagat cgaggtcgag actctcgtcg aagacgaagc
     4141 cgaggccgga gccgtgccag cggccgccgt gagcaagggc gaggaggata acatggccat
     4201 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     4261 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     4321 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     4381 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     4441 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     4501 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     4561 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     4621 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     4681 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     4741 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     4801 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     4861 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     4921 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4981 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5041 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5101 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     5161 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5221 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5281 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5341 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5401 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5461 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5521 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5581 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5641 acattcaaat atgtatccg
//
