LOCUS       sequence-13256-e        7246 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 7246)
  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 7246)
  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..7246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     promoter        498..775
                     /label=chicken beta-actin promoter
     primer_bind     1055..1077
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1205..1214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1211..2137
                     /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             1211..2137
                     /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             2150..3577
                     /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
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     misc_feature    3640..4228
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3693..3713)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4111..4122)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4271..4392
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4282..4301)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4336..4355
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4554..4683
                     /label=AAV2 ITR
     primer_bind     complement(4702..4719)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4702..4718)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4711..4733)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4860..5315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4947..4966)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5157..5178
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5341..5445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5446..5514,5515..6306)
                     /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(5664..5683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6477..7065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6966..6985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatcgtacca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      301 ccattgacgt caatgggtgg actatttacg gtaaactgcc cacttggcag tacatcaagt
      361 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      421 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      481 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
      541 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
      601 ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
      661 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      721 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      781 cgctgcgacg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      841 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      901 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      961 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
     1021 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1081 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1141 aacgtgctgg ttgttgtgct gtctcatcat tttggcaaag aattcggtac cgcgggcccc
     1201 tagcgccacc atggactatg gcggcgcttt gtctgccgtc ggacgcgaac ttttgttcgt
     1261 tactaatcct gtggtggtga acgggtccgt cctggtccct gaggatcaat gttactgtgc
     1321 cggatggatt gaatctcgcg gcacgaacgg cgctcagacc gcgtcaaatg tcctgcagtg
     1381 gcttgcagca ggattcagca ttttgctgct gatgttctat gcctaccaaa cctggaaatc
     1441 tacatgcggc tgggaggaga tctatgtgtg cgccattgaa atggttaagg tgattctcga
     1501 gttctttttt gagtttaaga atccctctat gctctacctt gccacaggac accgggtgca
     1561 gtggctgcgc tatgcagagt ggctgctcac ttgtcctgtc atccttatcc acctgagcaa
     1621 cctcaccggc ctgagcaacg actacagcag gagaaccatg ggactccttg tctcagacat
     1681 cgggactatc gtgtgggggg ctaccagcgc catggcaacc ggctatgtta aagtcatctt
     1741 cttttgtctt ggattgtgct atggcgcgaa cacatttttt cacgccgcca aagcatatat
     1801 cgagggttat catactgtgc caaagggtcg gtgccgccag gtcgtgaccg gcatggcatg
     1861 gctgtttttc gtgagctggg gtatgttccc aattctcttc attttggggc ccgaaggttt
     1921 tggcgtcctg agcgtctatg gctccaccgt aggtcacacg attattgatc tgatgagtaa
     1981 aaattgttgg gggttgttgg gacactacct gcgcgtcctg atccacgagc acatattgat
     2041 tcacggagat atccgcaaaa ccaccaaact gaacatcggc ggaacggaga tcgaggtcga
     2101 gactctcgtc gaagacgaag ccgaggccgg agccgtgcca gccgcggccg tgagcaaggg
     2161 cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
     2221 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2281 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     2341 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
     2401 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
     2461 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     2521 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2581 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     2641 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     2701 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     2761 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     2821 cctggggcat ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga
     2881 caacaacatg gccgtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat
     2941 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     3001 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     3061 cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta
     3121 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     3181 cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt
     3241 gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
     3301 ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
     3361 ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat
     3421 ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac
     3481 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca
     3541 cctgttcctg tacggcatgg acgagctgta caagtaaggg gatccactcg agtggagctc
     3601 gcgactagtc gattcgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     3661 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3721 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3781 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3841 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3901 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3961 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4021 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     4081 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4141 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4201 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg gccgcttcga
     4261 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     4321 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4381 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     4441 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt
     4501 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     4561 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4621 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4681 cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4741 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4801 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     4861 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4921 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4981 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5041 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5101 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5161 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5221 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5281 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     5341 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5401 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5461 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5521 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5581 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5641 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5701 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5761 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5821 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5881 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5941 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6001 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6061 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6121 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6181 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6241 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6301 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6361 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6421 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6481 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6541 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6601 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6661 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6721 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6781 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6841 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6901 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6961 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7021 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7081 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7141 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7201 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
