LOCUS       sequence-444309-        5688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of dominant-negative thyroid receptor beta (THRB, human) 
            with an HA tag in neurons driven by human synapsin promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5688)
  AUTHORS   Hochbaum DR, Hulshof L, Urke A, Wang W, Dubinsky AC, Farnsworth HC, 
            Hakim R, Lin S, Kleinberg G, Robertson K, Park C, Solberg A, Yang Y,
            Baynard C, Nadaf NM, Beron CC, Girasole AE, Chantranupong L, 
            Cortopassi MD, Prouty S, Geistlinger L, Banks AS, Scanlan TS, Datta 
            SR, Greenberg ME, Boulting GL, Macosko EZ, Sabatini BL
  TITLE     Thyroid hormone remodels cortex to coordinate body-wide metabolism 
            and exploration.
  JOURNAL   Cell. 2024 Oct 3;187(20):5679-5697.e23. doi: 
            10.1016/j.cell.2024.07.041. Epub 2024 Aug 22.
  PUBMED    39178853
REFERENCE   2  (bases 1 to 5688)
  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: 
            "10.1016/j.cell.2024.07.041"; journalName: "Cell"; date: "2024-10-3-
            3"; volume: "187"; issue: "20"; pages: "5679-5697.e23"
COMMENT     Sequence Label: DRH016_ AAV-hSyn-DN-THR
FEATURES             Location/Qualifiers
     source          1..5688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(206..653)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   706..835
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(897..1485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(977..996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1656..2447,2448..2516))
                     /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     2279..2298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2517..2621)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2689..2707
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2745..2767)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2867..2886
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2903..3358)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3040..3061)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3252..3271
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3433..3573
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3444..3573
                     /label=AAV2 ITR
     polyA_signal    3691..3802
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3836..4424
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3942..3953
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4351..4371
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4443..4469)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 atgagatgtg gcgacgactg ttcatttttc aacgtgctgc gcctctctga atggctcttc
       61 ctatgtaggc aggcttcaga cattcctact agcttccagt cgtgttctcg gtctggacag
      121 tgcttcggtt tgtcccaggc tgtaaggcca ttttctgtca tactgttggg agtcatggtg
      181 gcggatcctc tagccggttt ctcgactgcg ctctcaggca cgacacgact cctccgctgc
      241 ccaccgcaga ctgaggcagc gctgagtcgc cggcgccgca gcgcagatgg tcgcgcccgt
      301 gcccccctat ctcgcgcctc gcgtggtgcg gtccggctgg gccggcggcg gcgcggacgc
      361 gaccaaggtg gccgggaagg ggagtttgcg ggggaccggc gagtgacgtc agcgcgcctt
      421 cagtgctgag gcggcggtgg cgcgcgccgc caggcggggg cgaaggcact gtccgcggtg
      481 ctgaagctgg cagtgcgcac gcgcctcgcc gcatcctgtt tcccctcccc ctctctgata
      541 ggggatgcgc aatttgggga atgggggttg ggtgcttgtc cagtgggtcg gggtcggtcg
      601 tcaggtaggc acccccaccc cgcctcatcc tggtcctaaa acccacttgc actcatacgc
      661 agggccctct gcagtctaga cacttaatta aacgcgtgcg gccgcaggaa cccctagtga
      721 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
      781 tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg
      841 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      901 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      961 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1021 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1081 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1141 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1201 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1261 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1321 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1381 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1441 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1501 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1561 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1621 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1681 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1741 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1801 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1861 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1921 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1981 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2041 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2101 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2161 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2221 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     2281 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2341 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2401 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2461 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2521 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2581 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2641 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2701 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2761 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2821 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggcgta actatgcggc atcagagcag
     2881 attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta
     2941 aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat
     3001 aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca
     3061 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3121 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
     3181 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3241 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg
     3301 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac
     3361 tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3421 gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     3481 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     3541 agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc
     3601 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     3661 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     3721 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     3781 ggctggcaac tagaaggcac agtcgaagat ctctcgagca gcgctcggta tcgatgcggg
     3841 gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc
     3901 cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg
     3961 tagcagaagg acgtcccgcg cagaatccag gtggcaacat aggcgagcag ccaaggaaag
     4021 gacgatgatt tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc
     4081 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg
     4141 aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca
     4201 gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga
     4261 gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
     4321 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     4381 ataccagtca atctttcaca aattttgtaa tccagaggtt gattatcgat aagcttttac
     4441 taagcgtaat ctggaacatc gtatgggtaa cttccgcctc caccatcctc gaacacttcc
     4501 aagaacaaag gggggaagag ttctgtgggg cattccacct tcatgtgcag gaagcggctg
     4561 gcatggcagg ctcctatcat ccgcagatct gtcaccttca tcaggagttt tggccaaaag
     4621 tgtgtcacgt ggtgttttcg gtaattgata tagtgttcaa aggccagcag gaaactatct
     4681 tggtactttt ctattctctc aacacaggca agccccgggc gatctgaaga catcagcagg
     4741 acggcctgaa ggagggctac ttcagtgtca tccaggttga aagaagacag agacatgccc
     4801 aggtcaaaga tggcgtctga caccacccca agacccccat ttttcagctg gcccctcact
     4861 gccatttccc cattcaaggt taaagtctca ctttctgggt catagcgcac agcagcgcga
     4921 agggacatga tctccatgca gcagcctttg aggaggatga tctggtcttc acatggcagc
     4981 tcacaaaaca taggcaactt tttggcaaaa tccaccactc tggtaattgc tggtgtgatg
     5041 atttttgtaa aatggctgaa ggcttccaag tcaacctttc caccttctgg ggcattgact
     5101 attggtgctt gtccaatgtc ttctggcagg aatttccgtt tttgcttcca gtggctgcct
     5161 tgggcgttgg tcgccacatg ggcttcggtg acagttttga tgagctccca ttcctcgtct
     5221 gtgggctctg gcttgtgccc gatggacttc tgcagctctt cccgccgtct tttctcccgg
     5281 ttctcctcta tcagcttcct cttggccagc ctcttgctgt catccagcac caaatctgtt
     5341 gccatgccaa catagatgca tttcttaaag cgacattcct ggcactgatt tcgcgtgact
     5401 ttgtctatga cacattttcc ttcatattta caggaatagg atggatggag atttttctga
     5461 atggttcttc taaagaaacc cttgcagcct tcacacgtga tacagcggta gtgatacccg
     5521 gtggctttgt caccacacac tacacagagc tcgtccttgt ctaagtaact ggggatgtac
     5581 cctttacatt tcttctcctc cgtttggaag gtctgggcac ttgagacact ctggtcgttc
     5641 acatcatcat ggtccagatg gaatattgag ctagtccaag tggtctgg
//
