LOCUS       sequence-209250-        5769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with hSynapsin promoter, Cre-ON ChETA-mRuby2 (for 
            optogenetic activation), and W3SL regulatory cassette (for maximize 
            cloning capacity).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5769)
  AUTHORS   Li SJ, Vaughan A, Sturgill JF, Kepecs A
  TITLE     A Viral Receptor Complementation Strategy to Overcome CAV-2 Tropism 
            for Efficient Retrograde Targeting of Neurons.
  JOURNAL   Neuron. 2018 Jun 6;98(5):905-917.e5. doi: 
            10.1016/j.neuron.2018.05.028.
  PUBMED    29879392
REFERENCE   2  (bases 1 to 5769)
  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"; doi: 
            "10.1016/j.neuron.2018.05"; journalName: "Neuron"; date: "2018-06-6-
            6"; volume: "98"; issue: "5"; pages: "905-917.e5"
COMMENT     Sequence Label: pAAV-hSyn-DIO {ChETA-mRuby2}on-W3SL
FEATURES             Location/Qualifiers
     source          1..5769
                     /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        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(671..704)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(755..788)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(801..1511)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     regulatory      1508..1517
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1527..2453)
                     /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
                     PSMLYLATGHRVQWLRYATWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2450..2459
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2464..2497
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2548..2581
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2651..2671)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3031..3171
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3031..3160
                     /label=AAV2 ITR
     rep_origin      3246..3701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3333..3352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3543..3564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3718..3737)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3837..3859
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3897..3915)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3983..4087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4088..4156,4157..4948)
                     /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(4306..4325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5119..5707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5608..5627
                     /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 cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
      721 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
      781 gaagttatgg cgcgtaatta cttgtacagc tcgtccatcc caccaccaag cccggcgaac
      841 ttggcaactg cgtgttcgcg ttgtactacg aacatttcat tgtcactttc ctctaacctt
      901 tccaggcggt gatcaacggc atggataccg ggcatcttga tgttcccgac ggtctttttt
      961 gacctgtaag ttgttacgaa agagcaagac agatggccac caccatcaac tttcagtgcc
     1021 atatgagtgt atcccctcag accaccatct gctggataca tcatctctgt attaggctcc
     1081 caacccttgg tcttcttctg catcacggga ccattggagg gaaagtttac ccctctgact
     1141 tggacgtggt aaacgagaca gccatcctca aggctggtgt cctgcatgac ggtgacgact
     1201 ccaccatctt cgtatctcgt aactctttcc caagtaaaac cctcaggaaa ggactgttta
     1261 aagaaatcag gaatgccttt cgggtacttg ataaaagtac ggctgccata catgaacgac
     1321 gtggcaagaa tgtcaaaggc aaatggcagg ggtcctccct cgatgacttt gatcctcatg
     1381 gtttgagttc ccatgtacgg attgccttct ccttcacctg tgcatttgaa ttggtggccg
     1441 ttgaccgaac cttccatgac caccttcata cgcatatttt ccttgatcag ctcttcgccc
     1501 ttagacacca tggtggcggc cgctggcacg gctccggcct cggcttcgtc ttcgacgaga
     1561 gtctcgacct cgatctccgt tccgccgatg ttcagtttgg tggttttgcg gatatctccg
     1621 tgaatcaata tgtgctcgtg gatcaggacg cgcaggtagt gtcccaacaa cccccaacaa
     1681 tttttactca tcagatcaat aatcgtgtga cctacggtgg agccatagac gctcaggacg
     1741 ccaaaacctt cgggccccaa aatgaagaga attgggaaca taccccagct cacgaaaaac
     1801 agccatgcca tgccggtcac gacctggcgg caccgaccct ttggcacagt atgataaccc
     1861 tcgatatatg ctttggcggc gtgaaaaaat gtgttcgcgc catagcacaa tccaagacaa
     1921 aagaagatga ctttaacata gccggttgcc atggcgctgg tagcccccca cacgatagtc
     1981 ccgatgtctg agacaaggag tcccatggtt ctcctgctgt agtcgttgct caggccggtg
     2041 aggttgctca ggcggataag gatgacagga caagtgagca gccaggttgc atagcgcagc
     2101 cactgcaccc ggtgtcctgt ggcaaggtag agcatagagg gattcttaaa ctcaaaaaag
     2161 aactcgagaa tcaccttaac catttcaatg gcgcacacat agatctcctc ccagccgcat
     2221 gtagatttcc aggtttggta ggcatagaac atcagcagca aaatgctgaa tcctgctgca
     2281 agccactgca ggacatttga cgcggtctga gcgccgttcg tgccgcgaga ttcaatccat
     2341 ccggcacagt aacattgatc ctcagggacc aggacggacc cgttcaccac cacaggatta
     2401 gtaacgaaca aaagttcgcg tccgacggca gacaaagcgc cgccatagtc catggtggct
     2461 agcataactt cgtataaagt atcctatacg aagttatttg ccttaaccca gaaattatca
     2521 ctgttattct ttagaatggt gcaaagaata acttcgtata atgtatgcta tacgaagtta
     2581 tgaattccgc tcgagataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2641 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2701 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     2761 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2821 tgggcactga caattccgtg gtgtttattt gtgaaatttg tgatgctatt gctttatttg
     2881 taaccatcta gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2941 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3001 gagatgtggg aggtttttta aagcggccgc aggaacccct agtgatggag ttggccactc
     3061 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3121 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     3181 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     3241 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3301 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3361 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3421 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     3481 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3541 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     3601 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     3661 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     3721 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3781 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3841 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3901 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3961 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4021 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4081 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4141 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4201 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4261 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4321 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4381 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4441 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4501 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4561 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4621 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4681 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4741 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4801 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4861 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4921 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4981 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5041 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5101 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5161 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5221 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5281 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5341 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5401 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5461 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5521 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5581 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5641 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5701 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5761 tcacatgtt
//
