LOCUS       sequence-333398-        6481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for orexin neuropeptides.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6481)
  AUTHORS   Duffet L, Kosar S, Panniello M, Viberti B, Bracey E, Zych AD, 
            Radoux-Mergault A, Zhou X, Dernic J, Ravotto L, Tsai YC, Figueiredo 
            M, Tyagarajan SK, Weber B, Stoeber M, Gogolla N, Schmidt MH, 
            Adamantidis AR, Fellin T, Burdakov D, Patriarchi T
  TITLE     A genetically encoded sensor for in vivo imaging of orexin 
            neuropeptides.
  JOURNAL   Nat Methods. 2022 Feb;19(2):231-241. doi: 
            10.1038/s41592-021-01390-2. Epub 2022 Feb 10.
  PUBMED    35145320
REFERENCE   2  (bases 1 to 6481)
  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: 
            "10.1038/s41592-021-01390-2"; journalName: "Nat Methods"; date: 
            "2022-02"; volume: "19"; issue: "2"; pages: "231-241"
COMMENT     Sequence Label: pAAV.hSynapsin1.OxLight1
FEATURES             Location/Qualifiers
     source          1..6481
                     /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)"
     intron          646..778
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     761..780
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2871..3459
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2924..2944)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3342..3353)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3477..3701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3744..3884
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3744..3873
                     /label=AAV2 ITR
     rep_origin      3959..4414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4046..4065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4256..4277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4431..4450)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4550..4572
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4610..4628)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4696..4800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4801..4869,4870..5661)
                     /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(5019..5038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5832..6420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6321..6340
                     /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 agtcgagatt aattaactcg agcaggtaag tatcaaggtt
      661 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
      721 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
      781 gctagcgttt aaacggtacc ggatccgtgc gcgccatgaa gacgatcatc gccctgagct
      841 acatcttctg cctggtgttc gccgactaca aggacgatga tgacgccatg tcaggcacaa
      901 aactggaaga ctcccctcct tgtcggaatt ggtcctccgc gtccgagctt aacgaaacgc
      961 aagaaccttt tcttaatccg acagactacg atgatgaaga gttccttagg tatctctgga
     1021 gggagtatct gcatcccaaa gaatatgaat gggtactgat cgcgggatat atcattgtct
     1081 ttgtggtggc cctgattggg aatgtactcg tatgtgtagc agtttggaaa aaccatcata
     1141 tgagaactgt taccaactat tttattgtaa acctgagcct tgcagacgtg ttggtgacaa
     1201 tcacgtgcct cccggctacc ctcgttgttg acataaccga gacatggttc ttcggtcaaa
     1261 gtctgtgtaa ggtgatacca tatctccaga ccgtgagcgt ttctgtgagc gtgcttactc
     1321 ttagctgtat agctctcgat cgctggtacg cgatatgtca ccctcacaag ttcaagagta
     1381 cggctaagag ggctaggaat tctatagtca ttatttggat cgtgtcatgc ataataatga
     1441 taccgcaggc gattgtcatg gaatgctcta cggtatttcc gggtttggca aataaaacga
     1501 cgttgtttac agtttgcgac gagagatggg gtggggagat ttatccgaag atgtatcaca
     1561 tatgtttctt cctcgtcact tatatggctc cgctctgctt gatggtcctg gcgtaccttc
     1621 aaatctttcg gaaactgtgg tgccgagagt tgtccagtct catcaatgtc tacataaagg
     1681 cggataaaca aaaaaacgga attaaagcaa atttcaagat aagacacaac attgaagacg
     1741 gtggcgttca gcttgcatat cattaccaac agaatacccc catcggcgac ggtccggtgc
     1801 ttctgccaga caaccattac ctgtccgtac aatcaaagtt gtccaaagac cctaatgaga
     1861 aacgcgacca tatggtactg cttgaattcg tgaccgcagc aggaataact ctgggcatgg
     1921 atgaactcta taagggagga acaggtggat caatggtatc aaagggcgag gaacttttca
     1981 caggagtcgt cccaattctt gttgagttgg acggcgacgt taatggacac aagttttccg
     2041 tcagtggcga gggagagggc gacgctacat atgggaagtt gacgctcaaa ttcatatgta
     2101 ctactggtaa gctgcctgta ccttggccga ctttggtcac taccttgacc tatggtgttc
     2161 agtgtttcag cagatacccg gaccatatga agcagcacga tttttttaaa agtgccatgc
     2221 ccgaaggtta tattcaagag cgcactatct tcttcaagga cgacgggaac tataagacgc
     2281 gggctgaggt aaagttcgag ggagatacat tggtcaatcg aattgagctt aaagggatcg
     2341 acttcaaaga agacggaaac atcttgggac acaagctgga gtataataat catgaccagt
     2401 tgagacagat tcgcgcgagg cggaaaacgg cacggatgct tatgatcgtt cttcttgtat
     2461 tcgctatatg ctatctccct atctccattc ttaacgtttt gaaaagagtg ttcggtatgt
     2521 tcgcacacac ggaagataga gaaactgtgt acgcgtggtt cactttcagc cattggctgg
     2581 tctacgccaa cagcgcagcg aatcctataa tctacaattt cctttcaggc aagtttcgag
     2641 aggaatttaa ggctgcgttt tcctgttgct gcctgggtgt tcaccatcgg caagaagatc
     2701 gactcacacg aggtcgaacg tcaactgaaa gcagaaagtc tcttacgaca caaatcagca
     2761 attttgataa cattagcaaa ctgtccgaac aggttgtctt gaccagcatc agtaccttgc
     2821 ccgccgcaaa cggcgccggc cctcttcaga attggtgaaa gcttatcgat aatcaacctc
     2881 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2941 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3001 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3061 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3121 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3181 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3241 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     3301 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3361 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3421 ctcagacgag tcggatctcc ctttgggccg cctccccgct ctcgagatac gtacgactgt
     3481 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3541 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3601 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3661 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagatc
     3721 tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     3781 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     3841 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     3901 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     3961 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4021 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4081 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4141 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     4201 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4261 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     4321 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4381 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     4441 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4501 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4561 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4621 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4681 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4741 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4801 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4861 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4921 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4981 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5041 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5101 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5161 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5221 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5281 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5341 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5401 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5461 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5521 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5581 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5641 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5701 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5761 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5821 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5881 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5941 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6001 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6061 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6121 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6181 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6241 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6301 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6361 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6421 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6481 t
//
