LOCUS       sequence-435310-        7609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of luciferase opsin fusion protein 
            (luminopsin) for bioluminescent optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7609)
  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 7609)
  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 EFIa DIO LMO11 (GeNL_SS-ChRmine)
FEATURES             Location/Qualifiers
     source          1..7609
                     /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      3536..3545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3845..3854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3879..3912
                     /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    3963..3996
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4021..4609
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4074..4094)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4492..4503)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4643..4754
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4872..5012
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4872..5001
                     /label=AAV2 ITR
     rep_origin      5087..5542
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5174..5193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5384..5405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5559..5578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5678..5700
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5738..5756)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5824..5928
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5929..5997,5998..6789)
                     /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(6147..6166)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6960..7548
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7449..7468
                     /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 ttcacacgtt gatgtcgatc tggtccaggg ggatgtactc gccctcgctg
     1621 gtgatcctgc tcttggcggc cgcagacagt ctccgcaggc gctcgccatt gctgctcagc
     1681 ctcatatcca gctccagctt ctggaactca tcctcgtcgt gtcccagctg ctccagcagc
     1741 cgaaacagct cctcatcata ggtctttctg aaccgggcca gggcaaagcc gtaacaggac
     1801 ttggccacga tgtcgcacac gcagtgggcc acctctgtcc agttatcgtc gatccagccg
     1861 aagcctctag ggcagatggc ccacaccagg ggaaacagaa tccacagcat ggagaaggta
     1921 aacacggcga acttcagcac gtgcaggctg tgctcggcgc ctgtatcctg ggccagctgg
     1981 gccagccggc tatactgctt ggccacgatg ctcatcacga tagagtaggc gaagataaac
     2041 cagaaacagc caaagccata ccaggcgtag gcgccattcc tcagcctagg gtcgccctct
     2101 gcgtagaact cggccagcac gccagacatc aggatggcaa agatgatggc gctacaagac
     2161 acgcgatagg gggccctcag ctgatacagc agatcgtaca ccagcattgg gcaggtcagc
     2221 atgtagtcgg catacctgta attgccgaag ccgatgatga aggggcccac atcgccgaag
     2281 ttgtggccca gtgtgtcggc cacgatcttg ccgaagtagc tcagcatggc aaagaaattc
     2341 acaaacaggg ccaggttctg ctcctcctga tagccggcgt aaaaggaggt tctcttgcta
     2401 gggccccgag atgtccagga ctcatagccc aggtagaatg tggtgattgt cagcacgata
     2461 aagcaagacc agtgggcgcc gatggccacg gagttcagct tggtatccag ataccagccg
     2521 tccattgtgc ccacgtagaa catctggtcg gtgcctggtg cgtgtgcggt ggcggatccg
     2581 gcaattccac cacactggac tagtgggtcg ccctcgaggt tagagatctt cttcagcaga
     2641 cggtaaccgg taacaccgtt gatggtaaca cggaacagca tagaaccttc aggggtgatc
     2701 agacgttcgt cgatgatctt gttaccgttc cacagggtac cggtaacggt gatcttatta
     2761 ccgtcgaaaa cagcgatacc ttcgtaagga cgaccgaagt agttcagcat gttaggggta
     2821 acaccgtcga taaccagggt accgtattcc atgataacct tgaaatgatg gtcgtcaaca
     2881 gggtaaacaa ccttgaaaac ttcttcgata tgagccatct ggtcagcaga cagaccttcg
     2941 taagggatga taacatggat gtcgatcttc agaccgtttt caccagaacg aacgatacgc
     3001 tggatagggg taacagaaac agccagggtc tgcagaacag aagaaacacc accctgttcc
     3061 agaacctggt ccaggttgta agcagcgatc tgttcccagt caccaacgaa gtcttcaaat
     3121 ccggtaaagg ccttttgcca ctccttgaag ttgagctcgg tcttggagtg cttgagctcc
     3181 gtcttacgga acacgtacat cggctggttc ttcagatagt tagccgccat tggcttggca
     3241 aaggtgtagg tggtccgcgc agtgctccgg tagcgcttgc catttccagt ggtgtaactc
     3301 cacttaaagg tactgatgat ggttttgtcg ttggggtaag tcttcttcga cctgcaccag
     3361 tccgcagcgg tcagcgagtt ggtcatcaca ggaccgtcag cagggaaacc agtccccttc
     3421 acctgggcct ctcctttgat gtggcttccc tcgtaggtgt agcggtagtt aacagtaagg
     3481 gaggcaccat cttcaaactg cattgtgcga tggacttggt atccggagcc atctaccatg
     3541 gcggcctgga aaggcgacat cccgtcaggg tagggcaggt actgatggaa gccatacccg
     3601 atatgaggga ccagaatcca gggggagaac tggaggtcac ccttggtgga cttcaggttt
     3661 aactcctcat aaccatcatt tggattgccg gtgccctgac ccaccatgtc aaagtccaca
     3721 ccgttgatgg agccaaagat gtgtaactca tgtgtcgctg ggaggctagc catgttatcc
     3781 tcctcgccct tgctcacggc ctcggccaca gcgatgcaga tcagggcaaa cagaactttg
     3841 actcccatgg tggcggatcc aagcttaccg gtgctagcat aacttcgtat aaagtatcct
     3901 atacgaagtt atttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa
     3961 gaataacttc gtataatgta tgctatacga agttatgaat tcgatatcaa gcttatcgat
     4021 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4081 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4141 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4201 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4261 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4321 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4381 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     4441 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4501 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4561 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     4621 gcgctgctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4681 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4741 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4801 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     4861 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4921 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4981 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     5041 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     5101 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5161 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5221 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5281 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     5341 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5401 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     5461 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5521 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     5581 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5641 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     5701 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     5761 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5821 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5881 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5941 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6001 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6061 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6121 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6181 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6241 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6301 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6361 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6421 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6481 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6541 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6601 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6661 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6721 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6781 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6841 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6901 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6961 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7021 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7081 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7141 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7201 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7261 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7321 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7381 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7441 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7501 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7561 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
