LOCUS       sequence-165417-        5660 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 5660)
  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 5660)
  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..5660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    609..665
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             679..1605
                     /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             1618..2325
                     /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(1760..1778)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2028..2047
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2448..2569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2485..2504)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2539..2558
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2576..3031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2713..2734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2925..2944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3058..3162
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3164..3521
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3164..3355
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3187..3207)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3372..3507
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3556..4350
                     /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             3556..4350
                     /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(3610..3629)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4220..4239
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4538..4557)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4582..4629
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4958..5546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5447..5466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccacccg gtgccaccat ggactatggc ggcgctttgt ctgccgtcgg acgcgaactt
      721 ttgttcgtta ctaatcctgt ggtggtgaac gggtccgtcc tggtccctga ggatcaatgt
      781 tactgtgccg gatggattga atctcgcggc acgaacggcg ctcagaccgc gtcaaatgtc
      841 ctgcagtggc ttgcagcagg attcagcatt ttgctgctga tgttctatgc ctaccaaacc
      901 tggaaatcta catgcggctg ggagcagatc tatgtgtgcg ccattagaat ggttaaggtg
      961 attctcgagt tcttttttag ctttaagaat ccctctatgc tctaccttgc cacaggacac
     1021 cgggtgcagt ggctgcgcta tgcagagtgg ctgctcactt gtcctgtcat ccttatccac
     1081 ctgagcaacc tcaccggcct gagcaacgac tacagcagga gaaccatggg actccttgtc
     1141 tcaaacatcg ggtgtatcgt gtggggggct accagcgcca tggcaaccgg ctatgttaaa
     1201 gtcatcttct tttgtcttgg attgtgctat ggcgcgaaca cattttttca cgccgccaaa
     1261 gcatatatcg agggttatca tactgtgcca aagggtcggt gccgccaggt cgtgaccggc
     1321 atggcatggc tgtttttcgt gagctggggt atgttcccaa ttctcttcat tttggggccc
     1381 gaaggttttg gcgtcctgag cgtctatggc tccaccgtag gtcacacgat tattgatctg
     1441 atgagtaaaa attgttgggg gttgttggga cactacctgc gcgtcctgat ccacgagcac
     1501 atattgattc acggagatat ccgcaaaacc accaaactga acatcggcgg aacggagatc
     1561 gaggtcgaga ctctcgtcga agacgaagcc gaggccggag ccgtgccagc ggccgccgtg
     1621 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1681 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1741 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1801 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1861 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1921 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1981 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     2041 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     2101 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     2161 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     2221 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     2281 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     2341 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2401 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2461 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2521 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     2581 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2641 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2701 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2761 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2821 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2881 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2941 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3001 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3061 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3121 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3181 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3241 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3301 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3361 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3421 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3481 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3541 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3601 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3661 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3721 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3781 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3841 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3901 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3961 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4021 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4081 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4141 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4201 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4261 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4321 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4381 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4441 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4501 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4561 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4621 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4681 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4741 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4801 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4861 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4921 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4981 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5041 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5101 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5161 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5221 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5281 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5341 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5401 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5461 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5521 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5581 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5641 accgtattac cgccatgcat
//
