LOCUS       sequence-82241-e        7826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chloride-conducting Channelrhodopsin with slow kinetics, 
            neuron-specific promoter, plus red fluorescent protein, excitatory 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7826)
  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 7826)
  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: pAAV CaMKIIa ChR2 E90R D156N T159C 2A tDimer
FEATURES             Location/Qualifiers
     source          1..7826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..1299
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1317..1326
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1323..2249
                     /codon_start=1
                     /product="N-terminal fragment of channelrhodopsin-2 from 
                     the alga Chlamydomonas reinhardtii (Nagel et al., 2003)"
                     /label=ChR2
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIRMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSNIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             2259..2312
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      2328..2337
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2332..2341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2334..3011
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             3054..3728
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     misc_feature    3756..4344
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3809..3829)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4227..4238)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4347..4363)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4348..4364)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4424..4900
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4623..4642)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4940..5080
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4940..5069
                     /label=AAV2 ITR
     rep_origin      5155..5610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5242..5261)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5452..5473
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5627..5646)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5746..5768
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5806..5824)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5892..5996
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5997..6065,6066..6857)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6215..6234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7028..7616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7517..7536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7678..7818
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7689..7818
                     /label=AAV2 ITR
ORIGIN
        1 acgcgtttaa cattatggcc ttaggtcact tcatctccat ggggttcttc ttctgatttt
       61 ctagaaaatg agatgggggt gcagagagct tcctcagtga cctgcccagg gtcacatcag
      121 aaatgtcaga gctagaactt gaactcagat tactaatctt aaattccatg ccttgggggc
      181 atgcaagtac gatatacaga aggagtgaac tcattagggc agatgaccaa tgagtttagg
      241 aaagaagagt ccagggcagg gtacatctac accacccgcc cagccctggg tgagtccagc
      301 cacgttcacc tcattatagt tgcctctctc cagtcctacc ttgacgggaa gcacaagcag
      361 aaactgggac aggagcccca ggagaccaaa tcttcatggt ccctctggga ggatgggtgg
      421 ggagagctgt ggcagaggcc tcaggagggg ccctgctgct cagtggtgac agataggggt
      481 gagaaagcag acagagtcat tccgtcagca ttctgggtct gtttggtact tcttctcacg
      541 ctaaggtggc ggtgtgatat gcacaatggc taaaaagcag ggagagctgg aaagaaacaa
      601 ggacagagac agaggccaag tcaaccagac caattcccag aggaagcaaa gaaaccatta
      661 cagagactac aagggggaag ggaaggagag atgaattagc ttcccctgta aaccttagaa
      721 cccagctgtt gccagggcaa cggggcaata cctgtctctt cagaggagat gaagttgcca
      781 gggtaactac atcctgtctt tctcaaggac catcccagaa tgtggcaccc actagccgtt
      841 accatagcaa ctgcctcttt gccccactta atcccatccc gtctgttaaa agggccctat
      901 agttggaggt gggggaggta ggaagagcga tgatcacttg tggactaagt ttgttcgcat
      961 ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact tgctcctggg
     1021 cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc aggttttaaa
     1081 gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca gggcgtttcg
     1141 gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt gagcagccac
     1201 agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt tgcactcagg
     1261 agcacgggca ggcgagtggc ccctagttct gggggcagct cgaattcaag ctgctagcca
     1321 ccatggatta tggaggcgcc ctgagtgccg ttgggcgcga gctgctattt gtaacgaacc
     1381 cagtagtcgt caatggctct gtacttgtgc ctgaggacca gtgttactgc gcgggctgga
     1441 ttgagtcgcg tggcacaaac ggtgcccaaa cggcgtcgaa cgtgctgcaa tggcttgctg
     1501 ctggcttctc catcctactg cttatgtttt acgcctacca aacatggaag tcaacctgcg
     1561 gctgggagga gatctatgtg tgcgctatca ggatggtcaa ggtgattctc gagttcttct
     1621 tcgagtttaa gaacccgtcc atgctgtatc tagccacagg ccaccgcgtc cagtggttgc
     1681 gttacgccga gtggcttctc acctgcccgg tcattctcat tcacctgtca aacctgacgg
     1741 gcttgtccaa cgactacagc aggcgcacca tgggtctgct tgtgtctaat attggctgta
     1801 ttgtgtgggg cgccacttcc gccatggcca ccggatacgt caaggtcatc ttcttctgcc
     1861 tgggtctgtg ttatggtgct aacacgttct ttcacgctgc caaggcctac atcgagggtt
     1921 accacaccgt gccgaagggc cggtgtcgcc aggtggtgac tggcatggct tggctcttct
     1981 tcgtatcatg gggtatgttc cccatcctgt tcatcctcgg ccccgagggc ttcggcgtcc
     2041 tgagcgtgta cggctccacc gtcggccaca ccatcattga cctgatgtcg aagaactgct
     2101 ggggtctgct cggccactac ctgcgcgtgc tgatccacga gcatatcctc atccacggcg
     2161 acattcgcaa gaccaccaaa ttgaacattg gtggcactga gattgaggtc gagacgctgg
     2221 tggaggacga ggccgaggct ggcgcggtac ccgcggccga gggcagagga agtcttctaa
     2281 catgcggtga cgtggaggag aatcccggcc ctgcaccggg atccgcggcc gccatggtgg
     2341 cctcctccga ggacgtcatc aaagagttca tgcgcttcaa ggtgcgcatg gagggctccg
     2401 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2461 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
     2521 ccccccagtt ccagtacggc tccaaggcgt acgtgaagca ccccgccgac atccccgact
     2581 acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     2641 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcacgctg atctacaagg
     2701 tgaagttccg cggcaccaac ttcccccccg acggccccgt aatgcagaag aagaccatgg
     2761 gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc
     2821 accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag accatctaca
     2881 tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacaccaag ctggacatca
     2941 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag ggccgccacc
     3001 acctgttcct ggggcatggc accggcagca ccggcagcgg cagctccggc accgcctcct
     3061 ccgaggacgt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
     3121 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     3181 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     3241 agttccagta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gactacaaga
     3301 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     3361 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaagt
     3421 tccgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     3481 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg
     3541 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca
     3601 agaagcccgt gcagctgccc ggctactact acgtggacac caagctggac atcacctccc
     3661 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt
     3721 tcctgtaggt cgacttcgag caaaagctta tcgataatca acctctggat tacaaaattt
     3781 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3841 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3901 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3961 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4021 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4081 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4141 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     4201 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4261 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4321 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagacgcg atcgcggcgc
     4381 gccggcctct ctggcctcga cctcgagtct agagatgatc tacgggtggc atccctgtga
     4441 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     4501 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     4561 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     4621 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     4681 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     4741 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     4801 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     4861 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     4921 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4981 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     5041 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     5101 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     5161 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5221 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5281 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5341 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5401 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5461 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     5521 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5581 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5641 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5701 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5761 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5821 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5881 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5941 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6001 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6061 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6121 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6181 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6241 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6301 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6361 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6421 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6481 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6541 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6601 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6661 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6721 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6781 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6841 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6901 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6961 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7021 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7081 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7141 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     7201 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7261 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7321 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7381 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7441 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7501 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7561 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7621 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
     7681 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     7741 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     7801 ccatcactag gggttcctgc ggccgc
//
