LOCUS       sequence-467450-        7498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase opsin fusion protein (luminopsin) for bioluminescent 
            optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7498)
  AUTHORS   Ikefuama EC, Slaviero AN, Silvagnoli AD, Crespo EL, Schalau R, Gott 
            M, Tree MO, Dunbar GL, Rossignol J, Hochgeschwender U
  TITLE     Presymptomatic targeted circuit manipulation for ameliorating 
            Huntington's disease pathogenesis.
  JOURNAL   iScience. 2025 Feb 13;28(3):112022. doi: 10.1016/j.isci.2025.112022.
            eCollection 2025 Mar 21.
  PUBMED    40092615
REFERENCE   2  (bases 1 to 7498)
  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: "iScience.";
            date: "2025-02-13"; volume: "28(3):112022. doi"; pages: " 
            10.1016/j.isci.2025.112022. eCollection 2025 Mar 21"
COMMENT     Sequence Label: pAAV Ef1a DIO GeNL_SS hGtACR1
FEATURES             Location/Qualifiers
     source          1..7498
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /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(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3425..3434
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3734..3743
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3768..3801
                     /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    3852..3885
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3910..4498
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3963..3983)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4381..4392)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4532..4643
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4761..4901
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4761..4890
                     /label=AAV2 ITR
     rep_origin      4976..5431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5063..5082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5273..5294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5448..5467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5567..5589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5627..5645)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5713..5817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5818..5886,5887..6678)
                     /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(6036..6055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6849..7437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7338..7357
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccacgcg ttcaggtctc gccggctctg gcgtcctcgt acctctcggt gttgcccttc
     1621 tccaggtcct gctcgtactc aggcatcagg gtgccgtcca cgttcctgcc cttcacgtaa
     1681 tccctgtccc acttgccgtt cagcaggccc caggttgtag cccacagcag gatgccgtag
     1741 gtgttcttgc acagagcgtc gcccagcagg tacagcacgc tgctggtgtt ctcgctcatg
     1801 atgcaggcgc cggtgctgga gaagctccac aggatagggt aggccatcca gctcacgtag
     1861 aacacgatcc tcatcagctt cagcctctcc accaccttca gggacatggg ggagccgatg
     1921 gtctggaaat cgtggatggt aatggcgaaa atggcgaagg ccaccaccat ctcgaagatg
     1981 aagaagatgc tgctgatgaa acagtacagc cacacgtgca gcgggtctag caccacggag
     2041 gcggtgatgc cgaacacggt gcagatgctg gaggcggcga tcatcatagg gttgaggggg
     2101 atgctcttgt acttcacgag ggccatattg ctcaccaggc ccagcatgat agggcaggtg
     2161 cacagccagc tggccattct ggcccagggc aggtacttgc cattggccac tctgatgtag
     2221 ccgtttccgg tgaaggccag ggaataggtg atcatctcgg tggtgggcag gtagatggct
     2281 tcccagggca ccttgggggc tctggtcacc atgaagaaca cctgacaggc gctcacgacg
     2341 gccatcacca gctggacgta cttgatgccg tccagggtga tcaggctctt ctccacgcag
     2401 aactggttct ccctgctcca ttcgccgtag atggcgggat cacaggtgat gctgctggtg
     2461 gcggatccgg caattccacc acactggact agtgggtcgc cctcgaggtt agagatcttc
     2521 ttcagcagac ggtaaccggt aacaccgttg atggtaacac ggaacagcat agaaccttca
     2581 ggggtgatca gacgttcgtc gatgatcttg ttaccgttcc acagggtacc ggtaacggtg
     2641 atcttattac cgtcgaaaac agcgatacct tcgtaaggac gaccgaagta gttcagcatg
     2701 ttaggggtaa caccgtcgat aaccagggta ccgtattcca tgataacctt gaaatgatgg
     2761 tcgtcaacag ggtaaacaac cttgaaaact tcttcgatat gagccatctg gtcagcagac
     2821 agaccttcgt aagggatgat aacatggatg tcgatcttca gaccgttttc accagaacga
     2881 acgatacgct ggataggggt aacagaaaca gccagggtct gcagaacaga agaaacacca
     2941 ccctgttcca gaacctggtc caggttgtaa gcagcgatct gttcccagtc accaacgaag
     3001 tcttcaaatc cggtaaaggc cttttgccac tccttgaagt tgagctcggt cttggagtgc
     3061 ttgagctccg tcttacggaa cacgtacatc ggctggttct tcagatagtt agccgccatt
     3121 ggcttggcaa aggtgtaggt ggtccgcgca gtgctccggt agcgcttgcc atttccagtg
     3181 gtgtaactcc acttaaaggt actgatgatg gttttgtcgt tggggtaagt cttcttcgac
     3241 ctgcaccagt ccgcagcggt cagcgagttg gtcatcacag gaccgtcagc agggaaacca
     3301 gtccccttca cctgggcctc tcctttgatg tggcttccct cgtaggtgta gcggtagtta
     3361 acagtaaggg aggcaccatc ttcaaactgc attgtgcgat ggacttggta tccggagcca
     3421 tctaccatgg cggcctggaa aggcgacatc ccgtcagggt agggcaggta ctgatggaag
     3481 ccatacccga tatgagggac cagaatccag ggggagaact ggaggtcacc cttggtggac
     3541 ttcaggttta actcctcata accatcattt ggattgccgg tgccctgacc caccatgtca
     3601 aagtccacac cgttgatgga gccaaagatg tgtaactcat gtgtcgctgg gaggctagcc
     3661 atgttatcct cctcgccctt gctcacggcc tcggccacag cgatgcagat cagggcaaac
     3721 agaactttga ctcccatggt ggcggatcca agcttaccgg tgctagcata acttcgtata
     3781 aagtatccta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     3841 tggtgcaaag aataacttcg tataatgtat gctatacgaa gttatgaatt cgatatcaag
     3901 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3961 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4021 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4081 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4141 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4201 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4261 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     4321 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4381 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4441 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     4501 cgataccgag cgctgctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     4561 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4621 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4681 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     4741 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     4801 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4861 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     4921 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4981 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5041 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5101 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5161 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     5221 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5281 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     5341 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5401 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     5461 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5521 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5581 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5641 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5701 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5761 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5821 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5881 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5941 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6001 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6061 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6121 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6181 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6241 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6301 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6361 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6421 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6481 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6541 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6601 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6661 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6721 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6781 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6841 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6901 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6961 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7021 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7081 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7141 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7201 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7261 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7321 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7381 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7441 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
