LOCUS       sequence-170778-        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-ChloC
FEATURES             Location/Qualifiers
     source          1..5659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        51..408
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        51..242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(74..94)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      259..394
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             443..1237
                     /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             443..1237
                     /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(497..516)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1107..1126
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1425..1444)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1469..1516
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1845..2433
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2334..2353
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2608..2911
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2912..3115
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3066..3086
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3138..3218
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      3219..3228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3225..4151
                     /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
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             4164..4871
                     /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(4306..4324)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4574..4593
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4994..5115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5031..5050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5085..5104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5122..5577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5259..5280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5471..5490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5604..5659,1..49)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
       61 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      241 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
      301 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
      361 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
      421 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
      481 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
      541 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
      601 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
      661 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
      721 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
      781 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
      841 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
      901 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
      961 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     1021 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     1081 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     1141 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     1201 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     1261 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     1321 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     1381 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     1441 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     1501 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     1561 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     1621 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     1681 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1741 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1801 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1861 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1921 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1981 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2041 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2101 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2161 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2221 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2281 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2341 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2401 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2461 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2521 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     2581 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2641 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2701 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2761 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2821 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2881 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2941 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     3001 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     3061 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     3121 agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagctcaa gcttcgaatt
     3181 ctgcagtcga cggtaccgcg ggcccgggat ccaccggtgc caccatggac tatggcggcg
     3241 ctttgtctgc cgtcggacgc gaacttttgt tcgttactaa tcctgtggtg gtgaacgggt
     3301 ccgtcctggt ccctgaggat caatgttact gtgccggatg gattgaatct cgcggcacga
     3361 acggcgctca gaccgcgtca aatgtcctgc agtggcttgc agcaggattc agcattttgc
     3421 tgctgatgtt ctatgcctac caaacctgga aatctacatg cggctgggag gagatctatg
     3481 tgtgcgccat tagaatggtt aaggtgattc tcgagttctt ttttgagttt aagaatccct
     3541 ctatgctcta ccttgccaca ggacaccggg tgcagtggct gcgctatgca gagtggctgc
     3601 tcacttgtcc tgtcatcctt atccacctga gcaacctcac cggcctgagc aacgactaca
     3661 gcaggagaac catgggactc cttgtctcag acatcgggtg tatcgtgtgg ggggctacca
     3721 gcgccatggc aaccggctat gttaaagtca tcttcttttg tcttggattg tgctatggcg
     3781 cgaacacatt ttttcacgcc gccaaagcat atatcgaggg ttatcatact gtgccaaagg
     3841 gtcggtgccg ccaggtcgtg accggcatgg catggctgtt tttcgtgagc tggggtatgt
     3901 tcccaattct cttcattttg gggcccgaag gttttggcgt cctgagcgtc tatggctcca
     3961 ccgtaggtca cacgattatt gatctgatga gtaaaaattg ttgggggttg ttgggacact
     4021 acctgcgcgt cctgatccac gagcacatat tgattcacgg agatatccgc aaaaccacca
     4081 aactgaacat cggcggaacg gagatcgagg tcgagactct cgtcgaagac gaagccgagg
     4141 ccggagccgt gccagcggcc gccgtgagca agggcgagga ggataacatg gccatcatca
     4201 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     4261 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     4321 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     4381 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     4441 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     4501 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     4561 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     4621 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     4681 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     4741 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     4801 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     4861 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     4921 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4981 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5041 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5101 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5161 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5221 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5281 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5341 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5401 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5461 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5521 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     5581 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5641 caaatatgta tccgctcat
//
