LOCUS       sequence-450303-        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"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   826..955
                     /label=AAV2 ITR
     polyA_signal    1117..1238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1154..1173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1208..1227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    1281..1869
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1387..1398
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1796..1816
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1932..3359)
                     /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"
     CDS             complement(3372..4298)
                     /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             complement(3372..4298)
                     /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"
     regulatory      4295..4304
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4351..4370)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4432..4454)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4733..5011)
                     /label=chicken beta-actin promoter
     repeat_region   5324..5453
                     /label=AAV2 ITR
     rep_origin      5525..5980
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5612..5631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5822..5843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6006..6110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6111..6179,6180..6971)
                     /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(6329..6348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg cgctcgctcg
      841 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      901 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      961 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatccttatc
     1021 gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca tctccccctg
     1081 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg
     1261 cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac tcgtctgagg
     1321 gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc
     1381 cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc
     1441 aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc
     1501 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
     1561 cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat
     1621 gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac
     1681 aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat
     1741 gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag
     1801 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
     1861 aggttgatta tcgataagct tgatatcgaa ttcgaatcga ctagtcgcga gctccactcg
     1921 agtggatccc cttacttgta cagctcgtcc atgccgtaca ggaacaggtg gtggcggccc
     1981 tcggagcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc
     2041 ttggtgtcca cgtagtagta gccgggcagt tgcacgggct tcttggccat gtagatggtc
     2101 ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg gatctcgccc
     2161 ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc catggtcttc
     2221 ttctgcatta cggggccgtc gggggggaag ttggtgccgc gcatcttcac cttgtagatc
     2281 agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc gtcctcgaag
     2341 ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcttgta atcggggatg
     2401 tcggcggggt gcttcacgta cgccttggag ccgtacatga actgggggga caggatgtcc
     2461 caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg
     2521 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcat ggagccctcc
     2581 atgcgcacct tgaagcgcat gaactctttg atgacggcca tgttgttgtc ctcggaggag
     2641 gcggtgccgg agctgccgct gccggtgctg ccgatgccat gccccaggaa caggtggtgg
     2701 cggccctcgg agcgctcgta ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg
     2761 tccagcttgg tgtccacgta gtagtagccg ggcagttgca cgggcttctt ggccatgtag
     2821 atggtcttga actccaccag gtagtggccg ccgtccttca gcttcagggc ctggtggatc
     2881 tcgcccttca gcacgccgtc gcgggggtac aggcgctcgg tggaggcctc ccagcccatg
     2941 gtcttcttct gcattacggg gccgtcgggg gggaagttgg tgccgcgcat cttcaccttg
     3001 tagatcagcg tgccgtcctg cagggaggag tcctgggtca cggtcaccag accgccgtcc
     3061 tcgaagttca tcacgcgctc ccacttgaag ccctcgggga aggacagctt cttgtaatcg
     3121 gggatgtcgg cggggtgctt cacgtacgcc ttggagccgt acatgaactg gggggacagg
     3181 atgtcccagg cgaagggcag ggggccgccc ttggtcacct tcagcttggc ggtctgggtg
     3241 ccctcgtagg ggcggccctc gccctcgccc tcgatctcga actcgtggcc gttcatggag
     3301 ccctccatgc gcaccttgaa gcgcatgaac tctttgatga cctcctcgcc cttgctcacg
     3361 gccgcggctg gcacggctcc ggcctcggct tcgtcttcga cgagagtctc gacctcgatc
     3421 tccgttccgc cgatgttcag tttggtggtt ttgcggatat ctccgtgaat caatatgtgc
     3481 tcgtggatca ggacgcgcag gtagtgtccc aacaaccccc aacaattttt actcatcaga
     3541 tcaataatcg tgtgacctac ggtggagcca tagacgctca ggacgccaaa accttcgggc
     3601 cccaaaatga agagaattgg gaacataccc cagctcacga aaaacagcca tgccatgccg
     3661 gtcacgacct ggcggcaccg accctttggc acagtatgat aaccctcgat atatgctttg
     3721 gcggcgtgaa aaaatgtgtt cgcgccatag cacaatccaa gacaaaagaa gatgacttta
     3781 acatagccgg ttgccatggc gctggtagcc ccccacacga tagtcccgat gtctgagaca
     3841 aggagtccca tggttctcct gctgtagtcg ttgctcaggc cggtgaggtt gctcaggtgg
     3901 ataaggatga caggacaagt gagcagccac tctgcatagc gcagccactg cacccggtgt
     3961 cctgtggcaa ggtagagcat agagggattc ttaaactcaa aaaagaactc gagaatcacc
     4021 ttaaccattt caatggcgca cacatagatc tcctcccagc cgcatgtaga tttccaggtt
     4081 tggtaggcat agaacatcag cagcaaaatg ctgaatcctg ctgcaagcca ctgcaggaca
     4141 tttgacgcgg tctgagcgcc gttcgtgccg cgagattcaa tccatccggc acagtaacat
     4201 tgatcctcag ggaccaggac ggacccgttc accaccacag gattagtaac gaacaaaagt
     4261 tcgcgtccga cggcagacaa agcgccgcca tagtccatgg tggcgctagg ggcccgcggt
     4321 accgaattct ttgccaaaat gatgagacag cacaataacc agcacgttgc ccaggagctg
     4381 taggaaaaag aagaaggcat gaacatggtt agcagaggct ctagagccgc cggtcacacg
     4441 ccagaagccg aaccccgccc tgccccgtcc cccccgaagg cagccgtccc cccgcggaca
     4501 gccccgagcc gctccccccg cacaaagggc cctcccggag cccctcaagg ctttcacgca
     4561 gccacagaaa agaaacaagc cgtcattaaa ccaagcgcta attacagccc ggaggagaag
     4621 ggccgtcccg cccgctcacc tgtgggagta acgcggtcag tcagagccgg ggcgggcggc
     4681 gcgaggcggc ggcggagcgg ggcacggggc gaaggcagcg cgcagcgact cccgcccgcc
     4741 gcgcgcttcg ctttttatag ggccgccgcc gccgccgcct cgccataaaa ggaaactttc
     4801 ggagcgcgcc gctctgattg gctgccgccg cacctctccg cctcgccccg ccccgcccct
     4861 cgccccgccc cgccccgcct ggcgcgcgcc cccccccccc ccccgccccc atcgctgcac
     4921 aaaataatta aaaaataaat aaatacaaaa ttgggggtgg ggaggggggg ggagatgggg
     4981 agagtgaagc agaacgtggg gctcacctcg accatggtaa tagcgatgac taatacgtag
     5041 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     5101 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     5161 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     5221 ggcgttacta tgggaacata cgtcattatt gacgtcaatg gtacgatctt cctagagcat
     5281 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     5341 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     5401 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt
     5461 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5521 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5581 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5641 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5701 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5761 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5821 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5881 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5941 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     6001 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6061 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6121 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6181 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6241 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6301 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6361 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6421 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6481 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6541 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6601 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6661 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6721 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6781 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6841 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6901 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6961 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7021 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7081 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7141 c
//
