LOCUS       sequence-170775-        5659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Historical plasmid. For inhibition of neuronal activity, #66709 is a
            light-gated chloride channel with improved anion selectivity 
            (iChloC).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5659)
  AUTHORS   Wietek J, Wiegert JS, Adeishvili N, Schneider F, Watanabe H, Tsunoda
            SP, Vogt A, Elstner M, Oertner TG, Hegemann P
  TITLE     Conversion of Channelrhodopsin into a Light-Gated Chloride Channel.
  JOURNAL   Science. 2014 Mar 27.
  PUBMED    24674867
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: "Science. 
            2014 Mar 27."
COMMENT     Sequence Label: p-mCherry-N1-slowChloC
FEATURES             Location/Qualifiers
     source          1..5659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        159..350
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     307..327
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(352..456)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      483..938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(570..589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     780..801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    945..1066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(956..975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1010..1029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1189..1896)
                     /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(1467..1486)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1736..1754
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1909..2835)
                     /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
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIRMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSNIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2832..2841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2842..2922
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(2945..3148)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2974..2994)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3149..3452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3627..4215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3707..3726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4544..4591
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4616..4635
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4823..5617)
                     /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             complement(4823..5617)
                     /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(4934..4953)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5544..5563
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5652..5659,1..350))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
       61 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      301 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
      361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      481 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      541 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      661 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      781 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      841 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      901 taacgcgaat tttaacaaaa tattaacgct tacaatttac gccttaagat acattgatga
      961 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1021 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1081 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1141 cctctacaaa tgtggtatgg ctgattatga tctagagtcg cggccgcttt acttgtacag
     1201 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt
     1261 gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag
     1321 ctgcacgggc ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc
     1381 cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg
     1441 ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa
     1501 gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg
     1561 ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc
     1621 ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga
     1681 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     1741 caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat
     1801 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     1861 gatgatggcc atgttatcct cctcgccctt gctcacggcg gccgctggca cggctccggc
     1921 ctcggcttcg tcttcgacga gagtctcgac ctcgatctcc gttccgccga tgttcagttt
     1981 ggtggttttg cggatatctc cgtgaatcaa tatgtgctcg tggatcagga cgcgcaggta
     2041 gtgtcccaac aacccccaac aatttttact catcagatca ataatcgtgt gacctacggt
     2101 ggagccatag acgctcagga cgccaaaacc ttcgggcccc aaaatgaaga gaattgggaa
     2161 cataccccag ctcacgaaaa acagccatgc catgccggtc acgacctggc ggcaccgacc
     2221 ctttggcaca gtatgataac cctcgatata tgctttggcg gcgtgaaaaa atgtgttcgc
     2281 gccatagcac aatccaagac aaaagaagat gactttaaca tagccggttg ccatggcgct
     2341 ggtagccccc cacacgatac acccgatgtt tgagacaagg agtcccatgg ttctcctgct
     2401 gtagtcgttg ctcaggccgg tgaggttgct caggtggata aggatgacag gacaagtgag
     2461 cagccactct gcatagcgca gccactgcac ccggtgtcct gtggcaaggt agagcataga
     2521 gggattctta aactcaaaaa agaactcgag aatcacctta accattctaa tggcgcacac
     2581 atagatctcc tcccagccgc atgtagattt ccaggtttgg taggcataga acatcagcag
     2641 caaaatgctg aatcctgctg caagccactg caggacattt gacgcggtct gagcgccgtt
     2701 cgtgccgcga gattcaatcc atccggcaca gtaacattga tcctcaggga ccaggacgga
     2761 cccgttcacc accacaggat tagtaacgaa caaaagttcg cgtccgacgg cagacaaagc
     2821 gccgccatag tccatggtgg caccggtgga tcccgggccc gcggtaccgt cgactgcaga
     2881 attcgaagct tgagctcgag atctgagtcc ggtagcgcta gcggatctga cggttcacta
     2941 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     3001 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     3061 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     3121 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     3181 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     3241 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     3301 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     3361 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     3421 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     3481 atgaccccgt aattgattac tattaataac taatgcatgg cggtaatacg gttatccaca
     3541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct
     4381 atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg
     4441 gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg
     4501 gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca
     4561 ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc
     4621 tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc
     4681 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     4741 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     4801 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     4861 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     4921 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     4981 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     5041 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     5101 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     5161 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     5221 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     5281 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     5341 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     5401 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     5461 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     5521 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     5581 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     5641 cttgatcgat ctttgcaaa
//
