LOCUS       sequence-450732-        7141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses humanized ChR2 fused to tdTomato for optogenetic 
            activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7141)
  AUTHORS   Mao T, Kusefoglu D, Hooks BM, Huber D, Petreanu L, Svoboda K
  TITLE     Long-Range Neuronal Circuits Underlying the Interaction between 
            Sensory and Motor Cortex.
  JOURNAL   Neuron. 2011 Oct 6;72(1):111-23.
  PUBMED    21982373
REFERENCE   2  (bases 1 to 7141)
  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: "Neuron."; 
            date: "2011-10-6"; volume: "72(1)"; pages: "111-23"
COMMENT     Sequence Label: AAV-CAG-hChR2_tdTomato
FEATURES             Location/Qualifiers
     source          1..7141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1689..1818
                     /label=AAV2 ITR
     promoter        2131..2409
                     /label=chicken beta-actin promoter
     primer_bind     2688..2710
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2772..2791
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2838..2847
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2844..3770
                     /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
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             2844..3770
                     /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
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             3783..5210
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     misc_feature    5273..5861
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5326..5346)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5744..5755)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5904..6025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5915..5934)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5969..5988
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6187..6316
                     /label=AAV2 ITR
     rep_origin      complement(6553..7141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6633..6652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     1741 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     1801 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
     1861 catgctctag gaagatcgta ccattgacgt caataatgac gtatgttccc atagtaacgc
     1921 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     1981 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2041 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2101 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     2161 tccccatctc cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     2221 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc
     2281 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     2341 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     2401 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     2461 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     2521 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg
     2581 ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg
     2641 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     2701 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     2761 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgg
     2821 taccgcgggc ccctagcgcc accatggact atggcggcgc tttgtctgcc gtcggacgcg
     2881 aacttttgtt cgttactaat cctgtggtgg tgaacgggtc cgtcctggtc cctgaggatc
     2941 aatgttactg tgccggatgg attgaatctc gcggcacgaa cggcgctcag accgcgtcaa
     3001 atgtcctgca gtggcttgca gcaggattca gcattttgct gctgatgttc tatgcctacc
     3061 aaacctggaa atctacatgc ggctgggagg agatctatgt gtgcgccatt gaaatggtta
     3121 aggtgattct cgagttcttt tttgagttta agaatccctc tatgctctac cttgccacag
     3181 gacaccgggt gcagtggctg cgctatgcag agtggctgct cacttgtcct gtcatcctta
     3241 tccacctgag caacctcacc ggcctgagca acgactacag caggagaacc atgggactcc
     3301 ttgtctcaga catcgggact atcgtgtggg gggctaccag cgccatggca accggctatg
     3361 ttaaagtcat cttcttttgt cttggattgt gctatggcgc gaacacattt tttcacgccg
     3421 ccaaagcata tatcgagggt tatcatactg tgccaaaggg tcggtgccgc caggtcgtga
     3481 ccggcatggc atggctgttt ttcgtgagct ggggtatgtt cccaattctc ttcattttgg
     3541 ggcccgaagg ttttggcgtc ctgagcgtct atggctccac cgtaggtcac acgattattg
     3601 atctgatgag taaaaattgt tgggggttgt tgggacacta cctgcgcgtc ctgatccacg
     3661 agcacatatt gattcacgga gatatccgca aaaccaccaa actgaacatc ggcggaacgg
     3721 agatcgaggt cgagactctc gtcgaagacg aagccgaggc cggagccgtg ccagccgcgg
     3781 ccgtgagcaa gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg
     3841 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     3901 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     3961 tcctgtcccc ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc
     4021 ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     4081 aggacggcgg tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct
     4141 acaaggtgaa gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga
     4201 ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     4261 agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca
     4321 tctacatggc caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg
     4381 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc
     4441 gccaccacct gttcctgggg catggcatcg gcagcaccgg cagcggcagc tccggcaccg
     4501 cctcctccga ggacaacaac atggccgtca tcaaagagtt catgcgcttc aaggtgcgca
     4561 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     4621 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     4681 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg
     4741 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     4801 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc
     4861 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
     4921 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     4981 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     5041 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca
     5101 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
     5161 agggccgcca ccacctgttc ctgtacggca tggacgagct gtacaagtaa ggggatccac
     5221 tcgagtggag ctcgcgacta gtcgattcga attcgatatc aagcttatcg ataatcaacc
     5281 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5341 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5401 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5461 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5521 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5581 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5641 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     5701 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5761 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     5821 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacccgg
     5881 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5941 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6001 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     6061 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     6121 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     6181 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     6241 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     6301 gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6361 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6421 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6481 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6541 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6601 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6661 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6721 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6781 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6841 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6901 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6961 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7021 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7081 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7141 a
//
