LOCUS       sequence-247207-        6434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent vector expressing an optimized mosquito OPN3 rhodopsin
            in-frame with mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6434)
  AUTHORS   Mahn M, Saraf-Sinik I, Patil P, Pulin M, Bitton E, Karalis N, 
            Bruentgens F, Palgi S, Gat A, Dine J, Wietek J, Davidi I, Levy R, 
            Litvin A, Zhou F, Sauter K, Soba P, Schmitz D, Luthi A, Rost BR, 
            Wiegert JS, Yizhar O
  TITLE     Efficient optogenetic silencing of neurotransmitter release with a 
            mosquito rhodopsin.
  JOURNAL   Neuron. 2021 May 10. pii: S0896-6273(21)00161-6. doi: 
            10.1016/j.neuron.2021.03.013.
  PUBMED    33979634
REFERENCE   2  (bases 1 to 6434)
  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. 
            2021 May 10. pii: S0896-6273(21)00161-6. doi: 
            10.1016/j.neuron.2021.03.013."
COMMENT     Sequence Label: pAAV-hSyn1-SIO-eOPN3-mScarlet-WPRE
FEATURES             Location/Qualifiers
     source          1..6434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(636..669)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(703..1398)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(978..997)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1247..1265
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    2515..2548
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2561..3149
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2614..2634)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3032..3043)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3181..3657
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3380..3399)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3697..3837
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3697..3826
                     /label=AAV2 ITR
     rep_origin      3912..4367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3999..4018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4209..4230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4384..4403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4503..4525
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4563..4581)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4649..4753
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4754..4822,4823..5614)
                     /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(4972..4991)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5785..6373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6274..6293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtacctaccg ttcgtatagc atacattata
      661 cgaagttatg aattctagtt acacctcgtt ctcgtagcag aacttgtaca gctcgtccat
      721 gccgccggtg gagtggcggc cctcggagcg ttcgtactgt tccaccacgg tgtagtcctc
      781 gttgtgggag gtgatgtcca acttgcggtc gacgttgtag gcgccgggca tctgcacggg
      841 cttcttggcc ttgtaggtgg tcttgaagtc cgccaggtag cgtccgccgt ccttcaggcg
      901 cagggccatc ttaatgtcgc ccttcagcac gccgtcctcg gggtacaacc gctcggtgga
      961 cgcttcccag cccattgtct tcttctgcat tacggggccg tcaggaggga agttggtgcc
     1021 gcggagcttc accttgtaga tcagggtgcc gtcctccagg gaggtgtcct gggtcacggt
     1081 cacggcgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga
     1141 ctgcttatag tagtcgggga tgtcggcggg gtgcttggtg aaggccctgg agccgtacat
     1201 gaactgaggg gacaggatgt cccaggagaa gggcaggggg ccacccttgg tcaccttcag
     1261 cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc
     1321 gtggccgttc atggagccct ccatgtgcac cttgaaccgc atgaactcct tgatcactgc
     1381 ctcgcccttg ctcaccatca ccacgttgat gtcgatctgg tccaggggga tgtactcgcc
     1441 ctcgctggtg atcctgctct taccggttgg atctcgggct cgcggagcag gagccacctg
     1501 gctggtctcg gtcaccatgt ccctgctgct cttggtcagc tccctctgca tggtggtggt
     1561 gttctggtcg gcggtgggca cgccgccctt gttcctcacc ctggtgaagg cggccctgaa
     1621 ctgggtgttc atgcccacgt agatgatggg gttgtagcag atgctgctct tggcgatcag
     1681 agcaggcagc acagccagtc cagggccgat cagctcgggg gggccgaact gctcgatcag
     1741 ggcgaagatg gcgtaggggg tccaggccac catgaaggcc acgatcatca cggccaccat
     1801 gctggtcacc ctctgctcgg ccctgttgat cctgcccacc ctggcgctgt tcctcctcat
     1861 gtacacgatg atgttggtgt agctgtacac gatcacgatc aggggcacca ccaggccgaa
     1921 cacgaacagg aagatgatgt aggtggtggc gttcttggtc tggctctccc agttcacgct
     1981 gcagctgatg ttggcggcct cctgcacgta ggcgccccag ccgaacaggg gggggctggt
     2041 cagggcgaag ctgtagcccc agatgaagaa gatggccagg aaggcgccct tcctgctcag
     2101 gttcctgctg ctgaagggcc tgctgatcag gcagtacctc tcgtagctca gcacggtcag
     2161 ggtggtgatg ctggtgatgc ccagcaggct catgaagaag ccgtaggcca cgcacagggt
     2221 cctgccgaag atccacctgt ggctgatggc gctggtcagg gtcagggggt tgccgatgat
     2281 gctcacgctg aagtcgctgc acaccaggtt gaacaggatg atgttcatgg gggtccacag
     2341 ctgcatgtcc ttgctcatca gggcgatcac gaacaggttc aggaagaagc cgaagaagcc
     2401 gatgaagaac agggtgatgg cggcggcgtt gtaggcccag ggggccatca ggtcctcgta
     2461 gtcggccacg ctgctggcca cgtcgttggg ggcgtcgtac atggtggcgg atcctaccgt
     2521 tcgtataatg tatgctatac gaagttataa gcttatcgat aatcaacctc tggattacaa
     2581 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2641 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2701 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2761 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2821 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2881 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2941 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3001 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3061 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3121 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     3181 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3241 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3301 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     3361 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     3421 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     3481 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     3541 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     3601 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     3661 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3721 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3781 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3841 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3901 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3961 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     4261 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4381 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4441 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4501 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4561 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4621 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4681 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4741 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4801 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4861 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4921 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4981 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5041 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5101 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5161 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5221 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5281 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5341 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5401 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5461 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5521 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5581 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5641 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5701 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5761 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5821 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5881 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5941 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6001 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6061 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6121 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6181 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6241 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6301 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6361 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6421 ttttgctcac atgt
//
