LOCUS       sequence-444307-        5901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of dominant-negative thyroid receptor beta 
            (THRB, human) in neurons driven by nEF promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5901)
  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 5901)
  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: DRH032_ AAV-SIO-nEF-DN-THR
FEATURES             Location/Qualifiers
     source          1..5901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        188..399
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             412..680
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    complement(724..757)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(2156..2170,2171..2179))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      2179..2188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2189..2222
                     /label=loxKR3
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2247..2835
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2300..2320)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2718..2729)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3004..3085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3041..3060)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3095..3114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3164..3304
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3164..3293
                     /label=AAV2 ITR
     rep_origin      3379..3834
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3466..3485)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3676..3697
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3851..3870)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3970..3992
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4030..4048)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4116..4220
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4221..4289,4290..5081)
                     /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(4439..4458)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5252..5840
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5741..5760
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagga tctgcgatcg ctccggtgcc
      181 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
      241 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
      301 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
      361 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
      421 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
      481 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
      541 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
      601 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
      661 gttctgcgcc gttacagatc caagctgtga ccggcgccta cggatccaag tcactgcaga
      721 tgctaccgtt cgtatagcat acattatacg aagttattta ctaatcctcg aacacttcca
      781 agaacaaagg ggggaagagt tctgtggggc attccacctt catgtgcagg aagcggctgg
      841 catggcaggc tcctatcatc cgcagatctg tcaccttcat caggagtttt ggccaaaagt
      901 gtgtcacgtg gtgttttcgg taattgatat agtgttcaaa ggccagcagg aaactatctt
      961 ggtacttttc tattctctca acacaggcaa gccccgggcg atctgaagac atcagcagga
     1021 cggcctgaag gagggctact tcagtgtcat ccaggttgaa agaagacaga gacatgccca
     1081 ggtcaaagat ggcgtctgac accaccccaa gacccccatt tttcagctgg cccctcactg
     1141 ccatttcccc attcaaggtt aaagtctcac tttctgggtc atagcgcaca gcagcgcgaa
     1201 gggacatgat ctccatgcag cagcctttga ggaggatgat ctggtcttca catggcagct
     1261 cacaaaacat aggcaacttt ttggcaaaat ccaccactct ggtaattgct ggtgtgatga
     1321 tttttgtaaa atggctgaag gcttccaagt caacctttcc accttctggg gcattgacta
     1381 ttggtgcttg tccaatgtct tctggcagga atttccgttt ttgcttccag tggctgcctt
     1441 gggcgttggt cgccacatgg gcttcggtga cagttttgat gagctcccat tcctcgtctg
     1501 tgggctctgg cttgtgcccg atggacttct gcagctcttc ccgccgtctt ttctcccggt
     1561 tctcctctat cagcttcctc ttggccagcc tcttgctgtc atccagcacc aaatctgttg
     1621 ccatgccaac atagatgcat ttcttaaagc gacattcctg gcactgattt cgcgtgactt
     1681 tgtctatgac acattttcct tcatatttac aggaatagga tggatggaga tttttctgaa
     1741 tggttcttct aaagaaaccc ttgcagcctt cacacgtgat acagcggtag tgatacccgg
     1801 tggctttgtc accacacact acacagagct cgtccttgtc taagtaactg gggatgtacc
     1861 ctttacattt cttctcctcc gtttggaagg tctgggcact tgagacactc tggtcgttca
     1921 catcatcatg gtccagatgg aatattgagc tagtccaagt ggtctggatg agatgtggcg
     1981 acgactgttc atttttcaac gtgctgcgcc tctctgaatg gctcttccta tgtaggcagg
     2041 cttcagacat tcctactagc ttccagtcgt gttctcggtc tggacagtgc ttcggtttgt
     2101 cccaggctgt aaggccattt tctgtcatac tgttgggagt acttccgcct ccacccttat
     2161 cgtcatcgtc gtacagatcc atggtggcat aacaaggtat aatgtatgct atacgaagtt
     2221 atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2281 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2341 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2401 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2461 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2521 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2581 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2641 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     2701 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2761 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2821 gggccgcctc cccgcatcga taccgagcgc tgctcgagaa agtcactgca gatgccatca
     2881 agctggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2941 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3001 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3061 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3121 tcagcttatc gataccgcat gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     3181 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3241 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     3301 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3361 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3421 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     3481 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3541 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     3601 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3661 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     3721 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     3781 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     3841 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     3901 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3961 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4021 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4081 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4141 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4201 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4261 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4321 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4381 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4441 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4501 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4561 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4621 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4681 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4741 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4801 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4861 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4921 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4981 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5041 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5101 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5161 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5221 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5281 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5341 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5401 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5461 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5521 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5581 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5641 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5701 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5761 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5821 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5881 gctggccttt tgctcacatg t
//
