LOCUS       sequence-13257-e        7124 bp    DNA     circular SYN 23-APR-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 Apr 23, 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"
     repeat_region   56..185
                     /label=AAV2 ITR
     promoter        498..775
                     /label=chicken beta-actin promoter
     primer_bind     1054..1076
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1138..1157
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1234..1243
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1240..2166
                     /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"
     CDS             2179..3606
                     /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    3623..4211
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3676..3696)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4094..4105)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4254..4375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4265..4284)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4319..4338
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4537..4666
                     /label=AAV2 ITR
     rep_origin      4738..5193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4825..4844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5035..5056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5219..5323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5324..5392,5393..6184)
                     /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(5542..5561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6355..6943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6844..6863
                     /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 agtatttacg 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 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
      841 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      901 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
      961 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggctgt ccgcgggggg
     1021 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1081 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1141 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attggatcca ctcgagtgga
     1201 gctcgcgact agtcgattcg aattcgatct agcgccacca tggactatgg cggcgctttg
     1261 tctgccgtcg gacgcgaact tttgttcgtt actaatcctg tggtggtgaa cgggtccgtc
     1321 ctggtccctg aggatcaatg ttactgtgcc ggatggattg aatctcgcgg cacgaacggc
     1381 gctcagaccg cgtcaaatgt cctgcagtgg cttgcagcag gattcagcat tttgctgctg
     1441 atgttctatg cctaccaaac ctggaaatct acatgcggct gggaggagat ctatgtgtgc
     1501 gccattgaaa tggttaaggt gattctcgag ttcttttttg agtttaagaa tccctctatg
     1561 ctctaccttg ccacaggaca ccgggtgcag tggctgcgct atgcagagtg gctgctcact
     1621 tgtcctgtca tccttatccg cctgagcaac ctcaccggcc tgagcaacga ctacagcagg
     1681 agaaccatgg gactccttgt ctcagacatc gggactatcg tgtggggggc taccagcgcc
     1741 atggcaaccg gctatgttaa agtcatcttc ttttgtcttg gattgtgcta tggcgcgaac
     1801 acattttttc acgccgccaa agcatatatc gagggttatc atactgtgcc aaagggtcgg
     1861 tgccgccagg tcgtgaccgg catggcatgg ctgtttttcg tgagctgggg tatgttccca
     1921 attctcttca ttttggggcc cgaaggtttt ggcgtcctga gcgtctatgg ctccaccgta
     1981 ggtcacacga ttattgatct gatgagtaaa aattgttggg ggttgttggg acactacctg
     2041 cgcgtcctga tccacgagca catattgatt cacggagata tccgcaaaac caccaaactg
     2101 aacatcggcg gaacggagat cgaggtcgag actctcgtcg aagacgaagc cgaggccgga
     2161 gccgtgccag ccgcggccgt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc
     2221 aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag
     2281 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     2341 cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag
     2401 caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag
     2461 cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag
     2521 gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc
     2581 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac
     2641 ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg
     2701 gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac
     2761 gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2821 gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag caccggcagc
     2881 ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa agagttcatg
     2941 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     3001 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     3061 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac
     3121 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag
     3181 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc
     3241 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac
     3301 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
     3361 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac
     3421 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac
     3481 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     3541 cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac
     3601 aagtaagatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3661 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3721 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3781 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3841 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3901 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3961 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4021 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4081 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4141 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4201 cgcctccccg catcgatacc gtcgacccgg gcggccgctt cgagcagaca tgataagata
     4261 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     4321 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     4381 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     4441 caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg
     4501 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     4561 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4621 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgg cgtaatagcg
     4681 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     4741 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4801 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4861 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4921 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4981 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5041 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5101 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5161 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     5221 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5281 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5341 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5401 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5461 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5521 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5581 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5641 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5701 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5761 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5821 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5881 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5941 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6001 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6061 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6121 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6181 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6241 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6301 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6361 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6421 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6481 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6541 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6601 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6661 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6721 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6781 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6841 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6901 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6961 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7021 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7081 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
