LOCUS       sequence-444310-        5904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of wild-type 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 5904)
  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 5904)
  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: DRH031_ AAV-SIO-nEF-WT-THR
FEATURES             Location/Qualifiers
     source          1..5904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(52..66,67..75))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      75..84
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    85..118
                     /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    143..731
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(196..216)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(614..625)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    900..981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(937..956)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     991..1010
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1060..1200
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1060..1189
                     /label=AAV2 ITR
     rep_origin      1275..1730
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1362..1381)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1572..1593
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1747..1766)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1866..1888
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1926..1944)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2012..2116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2117..2185,2186..2977)
                     /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(2335..2354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3148..3736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3637..3656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3798..3938
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3809..3938
                     /label=AAV2 ITR
     promoter        3985..4196
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             4209..4477
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    complement(4521..4554)
                     /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)."
ORIGIN
        1 ggctgtaagg ccattttctg tcatactgtt gggagtactt ccgcctccac ccttatcgtc
       61 atcgtcgtac agatccatgg tggcataaca aggtataatg tatgctatac gaagttatga
      121 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
      181 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
      241 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
      301 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
      361 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
      421 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
      481 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
      541 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
      601 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
      661 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
      721 cgcctccccg catcgatacc gagcgctgct cgagagatct gtgatagcgg ccatcaagct
      781 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
      841 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
      901 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      961 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatcag
     1021 cttatcgata ccgcatgcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     1081 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     1141 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     1201 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     1261 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1321 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     1381 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1441 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     1501 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     1561 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     1621 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     1681 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     1741 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1801 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1861 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     1921 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1981 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2041 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2101 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2161 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2221 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2281 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2341 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2401 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2461 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2521 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2581 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2641 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2701 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2761 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2821 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2881 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2941 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3001 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3061 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3121 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3181 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3241 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3301 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3361 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3421 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3481 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3541 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3601 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3661 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3721 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3781 gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     3841 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     3901 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc gtaaggatct
     3961 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     4021 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
     4081 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     4141 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
     4201 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     4261 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     4321 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     4381 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     4441 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgg
     4501 atccgatctg tgatagcggc taccgttcgt atagcataca ttatacgaag ttatttacta
     4561 atcctcgaac acttccaaga acaaaggggg gaagagttct gtggggcatt ccaccttcat
     4621 gtgcaggaag cggctggcat ggcaggctcc tatcatccgc agatctgtca ccttcatcag
     4681 gagttttggc caaaagtgtg tcacgtggtg ttttcggtaa ttgatatagt gttcaaaggc
     4741 cagcaggaaa ctatcttggt acttttctat tctctcaaca caggcaagcc ccgggcgatc
     4801 tgaagacatc agcaggacgg cctgaaggag ggctacttca gtgtcatcca ggttgaaaga
     4861 agacagagac atgcccaggt caaagatggc gtctgacacc accccaagac ccccattttt
     4921 cagctggccc cgtgtcactg ccatttcccc attcaaggtt aaagtctcac tttctgggtc
     4981 atagcgcaca gcagcgcgaa gggacatgat ctccatgcag cagcctttga ggaggatgat
     5041 ctggtcttca catggcagct cacaaaacat aggcaacttt ttggcaaaat ccaccactct
     5101 ggtaattgct ggtgtgatga tttttgtaaa atggctgaag gcttccaagt caacctttcc
     5161 accttctggg gcattgacta ttggtgcttg tccaatgtct tctggcagga atttccgttt
     5221 ttgcttccag tggctgcctt gggcgttggt cgccacatgg gcttcggtga cagttttgat
     5281 gagctcccat tcctcgtctg tgggctctgg cttgtgcccg atggacttct gcagctcttc
     5341 ccgccgtctt ttctcccggt tctcctctat cagcttcctc ttggccagcc tcttgctgtc
     5401 atccagcacc aaatctgttg ccatgccaac atagatgcat ttcttaaagc gacattcctg
     5461 gcactgattt cgcgtgactt tgtctatgac acattttcct tcatatttac aggaatagga
     5521 tggatggaga tttttctgaa tggttcttct aaagaaaccc ttgcagcctt cacacgtgat
     5581 acagcggtag tgatacccgg tggctttgtc accacacact acacagagct cgtccttgtc
     5641 taagtaactg gggatgtacc ctttacattt cttctcctcc gtttggaagg tctgggcact
     5701 tgagacactc tggtcgttca catcatcatg gtccagatgg aatattgagc tagtccaagt
     5761 ggtctggatg agatgtggcg acgactgttc atttttcaac gtgctgcgcc tctctgaatg
     5821 gctcttccta tgtaggcagg cttcagacat tcctactagc ttccagtcgt gttctcggtc
     5881 tggacagtgc ttcggtttgt ccca
//
