LOCUS       sequence-333394-        6481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control non-binding 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.OxLight-ctr
FEATURES             Location/Qualifiers
     source          1..6481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        147..594
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          618..750
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     733..752
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2843..3431
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2896..2916)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3314..3325)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3449..3673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3716..3856
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3716..3845
                     /label=AAV2 ITR
     rep_origin      3931..4386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4018..4037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4228..4249
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4403..4422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4522..4544
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4582..4600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4668..4772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4773..4841,4842..5633)
                     /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(4991..5010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5804..6392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6293..6312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6454..6481,1..102)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
       61 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtgtgt
      121 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
      181 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
      241 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
      301 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
      361 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
      421 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
      481 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
      541 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga
      601 ttaattaact cgagcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
      661 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
      721 ctgacatcca ctttgccttt ctctccacag gtgctagcgt ttaaacggta ccggatccgt
      781 gcgcgccatg aagacgatca tcgccctgag ctacatcttc tgcctggtgt tcgccgacta
      841 caaggacgat gatgacgcca tgtcaggcac aaaactggaa gactcccctc cttgtcggaa
      901 ttggtcctcc gcgtccgagc ttaacgaaac gcaagaacct tttcttaatc cgacagacta
      961 cgatgatgaa gagttcctta ggtatctctg gagggagtat ctgcatccca aagaatataa
     1021 atgggtactg atcgcgggat atatcattgt ctttgtggtg gccctgattg ggaatgtact
     1081 cgtatgtgta gcagtttgga aaaaccatca tatgagaact gttaccaact attttattgt
     1141 aaacctgagc cttgcagacg tgttggtgac aatcacgtgc ctcccggctg ccctcgttgt
     1201 tgacataacc gagacatggt tcttcggtca aagtctgtgt aaggtgatac catatctcca
     1261 gaccgtgagc gtttctgtga gcgtgcttac tcttagctgt atagctctcg atcgctggta
     1321 cgcgatatgt caccctcaca agttcaagag tacggctaag agggctagga attctatagt
     1381 cattatttgg atcgtgtcat gcataataat gataccgcag gcgattgtca tggaatgctc
     1441 tacggtattt ccgggtttgg caaataaaac gacgttgttt acagtttgcg acgagagatg
     1501 gggtggggag atttatccga agatgtatca catatgtttc ttcctcgtca cttatatggc
     1561 tccgctctgc ttgatggtcc tggcgtacct tcaaatcttt cggaaactgt ggtgccgaga
     1621 gttgtccagt ctcatcaatg tctacataaa ggcggataaa caaaaaaacg gaattaaagc
     1681 aaatttcaag ataagacaca acattgaaga cggtggcgtt cagcttgcat atcattacca
     1741 acagaatacc cccatcggcg acggtccggt gcttctgcca gacaaccatt acctgtccgt
     1801 acaatcaaag ttgtccaaag accctaatga gaaacgcgac catatggtac tgcttgaatt
     1861 cgtgaccgca gcaggaataa ctctgggcat ggatgaactc tataagggag gaacaggtgg
     1921 atcaatggta tcaaagggcg aggaactttt cacaggagtc gtcccaattc ttgttgagtt
     1981 ggacggcgac gttaatggac acaagttttc cgtcagtggc gagggagagg gcgacgctac
     2041 atatgggaag ttgacgctca aattcatatg tactactggt aagctgcctg taccttggcc
     2101 gactttggtc actaccttga cctatggtgt tcagtgtttc agcagatacc cggaccatat
     2161 gaagcagcac gattttttta aaagtgccat gcccgaaggt tatattcaag agcgcactat
     2221 cttcttcaag gacgacggga actataagac gcgggctgag gtaaagttcg agggagatac
     2281 attggtcaat cgaattgagc ttaaagggat cgacttcaaa gaagacggaa acatcttggg
     2341 acacaagctg gagtataata atcatgacca gttgagacag attcgcgcga ggcggaaaac
     2401 ggcacggatg cttatgatcg ttcttcttgt attcgctata tgctatctcc ctatctccat
     2461 tcttaacgtt ttgaaaagag tgttcggtat gttcgcacac acggaagata gagaaactgt
     2521 gtacgcgtgg ttcactttca gccattggct ggtctacgcc aacagcgcag cgaatcctat
     2581 aatctacaat ttcctttcag gcaagtttcg agaggaattt aaggctgcgt tttcctgttg
     2641 ctgcctgggt gttcaccatc ggcaagaaga tcgactcaca cgaggtcgaa cgtcaactga
     2701 aagcagaaag tctcttacga cacaaatcag caattttgat aacattagca aactgtccga
     2761 acaggttgtc ttgaccagca tcagtacctt gcccgccgca aacggcgccg gccctcttca
     2821 gaattggtga aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2881 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2941 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3001 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3061 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3121 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3181 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3241 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3301 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3361 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3421 cgcctccccg ctctcgagat acgtacgact gtgccttcta gttgccagcc atctgttgtt
     3481 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3541 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3601 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3661 gtgggctcta tggcttctga ggcggaaaga tctcacgtgc ggaccgagcg gccgcaggaa
     3721 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3781 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3841 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3901 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3961 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4021 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4081 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4141 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4201 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4261 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4321 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4381 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4441 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4501 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4561 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4621 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4681 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4741 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4801 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4861 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4921 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4981 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5041 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5101 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5161 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5221 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5281 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5341 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5401 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5461 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5521 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5581 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5641 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5701 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5761 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5821 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5881 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5941 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6001 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6061 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6121 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6181 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6241 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6301 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6361 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6421 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc
     6481 t
//
