LOCUS       sequence-68465-e        7292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses channel rhodopsin 2 (H134R) (fused to 
            iRFP) under the CaM Kinase IIa promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7292)
  AUTHORS   Richie CT, Whitaker LR, Whitaker KW, Necarsulmer J, Baldwin HA, 
            Zhang Y, Fortuno L, Hinkle JJ, Koivula P, Henderson MJ, Sun W, Wang 
            K, Smith JC, Pickel J, Ji N, Hope BT, Harvey BK
  TITLE     Near-infrared fluorescent protein iRFP713 as a reporter protein for 
            optogenetic vectors, a transgenic Cre-reporter rat, and other 
            neuronal studies.
  JOURNAL   J Neurosci Methods. 2017 Jun 1;284:1-14. doi: 
            10.1016/j.jneumeth.2017.03.020. Epub 2017 Apr 2.
  PUBMED    28380331
REFERENCE   2  (bases 1 to 7292)
  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: "J Neurosci 
            Methods."; date: "2017-06-1"; volume: "284:1-14. doi"; pages: " 
            10.1016/j.jneumeth.2017.03.020. Epub 2017 Apr 2"
COMMENT     Sequence Label: pOTTC351 - pAAV CaMKIIa hChR2(H134R)-iRFP
FEATURES             Location/Qualifiers
     source          1..7292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        160..1448
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1487..1496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1493..2419
                     /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             2438..3388
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     misc_feature    3419..4007
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3472..3492)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3890..3901)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4039..4515
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4238..4257)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4555..4695
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4555..4684
                     /label=AAV2 ITR
     rep_origin      4770..5225
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4857..4876)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5067..5088
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5242..5261)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5361..5383
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5421..5439)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5507..5611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5612..5680,5681..6472)
                     /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(5830..5849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6643..7231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7132..7151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct taggtcactt
      181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
      241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
      301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
      361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
      421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
      481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
      541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
      601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
      661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
      721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
      781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
      841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
      901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
      961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
     1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
     1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
     1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
     1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
     1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
     1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
     1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
     1441 ggggcagctc tagagcggta ccggatcctc tagagtcgac tccggagcca ccatggacta
     1501 tggcggcgct ttgtctgccg tcggacgcga acttttgttc gttactaatc ctgtggtggt
     1561 gaacgggtcc gtcctggtcc ctgaggatca atgttactgt gccggatgga ttgaatctcg
     1621 cggcacgaac ggcgctcaga ccgcgtcaaa tgtcctgcag tggcttgcag caggattcag
     1681 cattttgctg ctgatgttct atgcctacca aacctggaaa tctacatgcg gctgggagga
     1741 gatctatgtg tgcgccattg aaatggttaa ggtgattctc gagttctttt ttgagtttaa
     1801 gaatccctct atgctctacc ttgccacagg acaccgggtg cagtggctgc gctatgcaga
     1861 gtggctgctc acttgtcctg tcatccttat ccgcctgagc aacctcaccg gcctgagcaa
     1921 cgactacagc aggagaacca tgggactcct tgtctcagac atcgggacta tcgtgtgggg
     1981 ggctaccagc gccatggcaa ccggctatgt taaagtcatc ttcttttgtc ttggattgtg
     2041 ctatggcgcg aacacatttt ttcacgccgc caaagcatat atcgagggtt atcatactgt
     2101 gccaaagggt cggtgccgcc aggtcgtgac cggcatggca tggctgtttt tcgtgagctg
     2161 gggtatgttc ccaattctct tcattttggg gcccgaaggt tttggcgtcc tgagcgtcta
     2221 tggctccacc gtaggtcaca cgattattga tctgatgagt aaaaattgtt gggggttgtt
     2281 gggacactac ctgcgcgtcc tgatccacga gcacatattg attcacggag atatccgcaa
     2341 aaccaccaaa ctgaacatcg gcggaacgga gatcgaggtc gagactctcg tcgaagacga
     2401 agccgaggcc ggagccgtgc cagcggccgc cactagtatg gctgaaggat ccgtcgccag
     2461 gcagcctgac ctcttgacct gcgacgatga gccgatccat atccccggtg ccatccaacc
     2521 gcatggactg ctgctcgccc tcgccgccga catgacgatc gttgccggca gcgacaacct
     2581 tcccgaactc accggactgg cgatcggcgc cctgatcggc cgctctgcgg ccgatgtctt
     2641 cgactcggag acgcacaacc gtctgacgat cgccttggcc gagcccgggg cggccgtcgg
     2701 agcaccgatc actgtcggct tcacgatgcg aaaggacgca ggcttcatcg gctcctggca
     2761 tcgccatgat cagctcatct tcctcgagct cgagcctccc cagcgggacg tcgccgagcc
     2821 gcaggcgttc ttccgccgca ccaacagcgc catccgccgc ctgcaggccg ccgaaacctt
     2881 ggaaagcgcc tgcgccgccg cggcgcaaga ggtgcggaag attaccggct tcgatcgggt
     2941 gatgatctat cgcttcgcct ccgacttcag cggcgaagtg atcgcagagg atcggtgcgc
     3001 cgaggtcgag tcaaaactag gcctgcacta tcctgcctca accgtgccgg cgcaggcccg
     3061 tcggctctat accatcaacc cggtacggat cattcccgat atcaattatc ggccggtgcc
     3121 ggtcacccca gacctcaatc cggtcaccgg gcggccgatt gatcttagct tcgccatcct
     3181 gcgcagcgtc tcgcccgtcc atctggaatt catgcgcaac ataggcatgc acggcacgat
     3241 gtcgatctcg attttgcgcg gcgagcgact gtggggattg atcgtttgcc atcaccgaac
     3301 gccgtactac gtcgatctcg atggccgcca agcctgcgag ctagtcgccc aggttctggc
     3361 ctggcagatc ggcgtgatgg aagagtgagt cgacgaattc gatatcaagc ttatcgataa
     3421 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3481 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3541 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3601 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3661 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3721 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3781 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     3841 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3901 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3961 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc
     4021 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     4081 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     4141 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     4201 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     4261 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     4321 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     4381 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     4441 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     4501 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac
     4561 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4621 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4681 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4741 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4801 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4861 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4921 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4981 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5041 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5101 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5161 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     5221 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5281 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5341 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5401 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5461 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5521 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5581 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5641 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5701 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5761 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5821 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5881 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5941 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6001 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6061 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6121 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6181 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6241 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6301 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6361 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6421 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6481 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6541 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6601 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6661 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6721 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6781 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6841 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6901 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6961 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7021 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7081 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7141 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7201 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7261 cctggccttt tgctggcctt ttgctcacat gt
//
