LOCUS       sequence-423533-        7124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of hChR2(H134R) for neuronal excitation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7124)
  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 7124)
  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-H134R-tdTomato
FEATURES             Location/Qualifiers
     source          1..7124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(104..123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     314..335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        498..602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(603..671,672..1463)
                     /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(821..840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1634..2222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2123..2142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     polyA_signal    2750..2871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2787..2806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2841..2860
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2914..3502
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3020..3031
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3429..3449
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3519..4946)
                     /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(4959..5885)
                     /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
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      5882..5891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5968..5987)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(6049..6071)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6350..6627)
                     /label=chicken beta-actin promoter
     repeat_region   6940..7069
                     /label=AAV2 ITR
ORIGIN
        1 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
       61 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
      121 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
      181 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
      241 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
      301 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
      361 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
      421 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
      481 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      541 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      601 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      661 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      721 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      781 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      841 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      901 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      961 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1021 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1081 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1141 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1201 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1261 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1321 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1381 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1441 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1501 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1561 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1621 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1681 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1741 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1801 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1861 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1921 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1981 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2041 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2101 agcgcacgag ggagcttccn rggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2161 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2221 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2281 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2341 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2401 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcctt atcgatttta ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2701 acatctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2761 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2821 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2881 ctcgaagcgg ccgcccgggt cgacggtatc gatgcgggga ggcggcccaa agggagatcc
     2941 gactcgtctg agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc
     3001 gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca
     3061 gaatccaggt ggcaacacag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca
     3121 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     3181 cgatgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac
     3241 aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc
     3301 cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca
     3361 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     3421 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     3481 ttttgtaatc cagaggttga ttatcgataa gcttgatctt acttgtacag ctcgtccatg
     3541 ccgtacagga acaggtggtg gcggccctcg gagcgctcgt actgttccac gatggtgtag
     3601 tcctcgttgt gggaggtgat gtccagcttg gtgtccacgt agtagtagcc gggcagttgc
     3661 acgggcttct tggccatgta gatggtcttg aactccacca ggtagtggcc gccgtccttc
     3721 agcttcaggg cctggtggat ctcgcccttc agcacgccgt cgcgggggta caggcgctcg
     3781 gtggaggcct cccagcccat ggtcttcttc tgcattacgg ggccgtcggg ggggaagttg
     3841 gtgccgcgca tcttcacctt gtagatcagc gtgccgtcct gcagggagga gtcctgggtc
     3901 acggtcacca gaccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg
     3961 aaggacagct tcttgtaatc ggggatgtcg gcggggtgct tcacgtacgc cttggagccg
     4021 tacatgaact ggggggacag gatgtcccag gcgaagggca gggggccgcc cttggtcacc
     4081 ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg
     4141 aactcgtggc cgttcatgga gccctccatg cgcaccttga agcgcatgaa ctctttgatg
     4201 acggccatgt tgttgtcctc ggaggaggcg gtgccggagc tgccgctgcc ggtgctgccg
     4261 atgccatgcc ccaggaacag gtggtggcgg ccctcggagc gctcgtactg ttccacgatg
     4321 gtgtagtcct cgttgtggga ggtgatgtcc agcttggtgt ccacgtagta gtagccgggc
     4381 agttgcacgg gcttcttggc catgtagatg gtcttgaact ccaccaggta gtggccgccg
     4441 tccttcagct tcagggcctg gtggatctcg cccttcagca cgccgtcgcg ggggtacagg
     4501 cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggggggg
     4561 aagttggtgc cgcgcatctt caccttgtag atcagcgtgc cgtcctgcag ggaggagtcc
     4621 tgggtcacgg tcaccagacc gccgtcctcg aagttcatca cgcgctccca cttgaagccc
     4681 tcggggaagg acagcttctt gtaatcgggg atgtcggcgg ggtgcttcac gtacgccttg
     4741 gagccgtaca tgaactgggg ggacaggatg tcccaggcga agggcagggg gccgcccttg
     4801 gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg
     4861 atctcgaact cgtggccgtt catggagccc tccatgcgca ccttgaagcg catgaactct
     4921 ttgatgacct cctcgccctt gctcacggcc gcggctggca cggctccggc ctcggcttcg
     4981 tcttcgacga gagtctcgac ctcgatctcc gttccgccga tgttcagttt ggtggttttg
     5041 cggatatctc cgtgaatcaa tatgtgctcg tggatcagga cgcgcaggta gtgtcccaac
     5101 aacccccaac aatttttact catcagatca ataatcgtgt gacctacggt ggagccatag
     5161 acgctcagga cgccaaaacc ttcgggcccc aaaatgaaga gaattgggaa cataccccag
     5221 ctcacgaaaa acagccatgc catgccggtc acgacctggc ggcaccgacc ctttggcaca
     5281 gtatgataac cctcgatata tgctttggcg gcgtgaaaaa atgtgttcgc gccatagcac
     5341 aatccaagac aaaagaagat gactttaaca tagccggttg ccatggcgct ggtagccccc
     5401 cacacgatag tcccgatgtc tgagacaagg agtcccatgg ttctcctgct gtagtcgttg
     5461 ctcaggccgg tgaggttgct caggcggata aggatgacag gacaagtgag cagccactct
     5521 gcatagcgca gccactgcac ccggtgtcct gtggcaaggt agagcataga gggattctta
     5581 aactcaaaaa agaactcgag aatcacctta accatttcaa tggcgcacac atagatctcc
     5641 tcccagccgc atgtagattt ccaggtttgg taggcataga acatcagcag caaaatgctg
     5701 aatcctgctg caagccactg caggacattt gacgcggtct gagcgccgtt cgtgccgcga
     5761 gattcaatcc atccggcaca gtaacattga tcctcaggga ccaggacgga cccgttcacc
     5821 accacaggat tagtaacgaa caaaagttcg cgtccgacgg cagacaaagc gccgccatag
     5881 tccatggtgg cgctagatcg aattcgaatc gactagtcgc gagctccact cgagtggatc
     5941 caattctttg ccaaaatgat gagacagcac aataaccagc acgttgccca ggagctgtag
     6001 gaaaaagaag aaggcatgaa catggttagc agaggctcta gagccgccgg tcacacgcca
     6061 gaagccgaac cccgccctgc cccgtncccc ccgaaggcag ccgtcccccc gcggacagcc
     6121 ccgagccgct ccccccgcac aaagggccct cccggagccc ctcaaggctt tcacgcagcc
     6181 acagaaaaga aacaagccgt cattaaacca agcgctaatt acagcccgga ggagaagggc
     6241 cgtcccgccc gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg
     6301 aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg
     6361 cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga
     6421 gcgcgccgct ctgattggct gccgccgcac ctctccgcct cgccccgccc cgcccctcgc
     6481 cccgccccgc cccgcctggc gcgcgncccc cccccccccc cgcccccatc gctgcacaaa
     6541 ataattaaaa aataaataaa tacaaaattg ggggtgggga ngggggggag atggggagag
     6601 tgaagcagaa cgtggggctc acctcgacca tggtaatagc gatgactaat acgtagatgt
     6661 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     6721 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     6781 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     6841 ttactatggg aacatacgtc attattgacg tcaatggtac gatcttccta gagcatggct
     6901 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     6961 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     7021 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tggcgtaata
     7081 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagc
//
