LOCUS       sequence-377905-        5850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adipocyte-specific overexpression of genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5850)
  AUTHORS   Xue K, Wu D, Wang Y, Zhao Y, Shen H, Yao J, Huang X, Li X, Zhou Z, 
            Wang Z, Qiu Y
  TITLE     The mitochondrial calcium uniporter engages UCP1 to form a 
            thermoporter that promotes thermogenesis.
  JOURNAL   Cell Metab. 2022 Sep 6;34(9):1325-1341.e6. doi: 
            10.1016/j.cmet.2022.07.011. Epub 2022 Aug 16.
  PUBMED    35977541
REFERENCE   2  (bases 1 to 5850)
  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.cmet.2022.07.011"; journalName: "Cell Metab"; date: 
            "2022-09-6- 6"; volume: "34"; issue: "9"; pages: "1325-1341.e6"
COMMENT     Sequence Label: pAAV-ADP-MCS-FLAG
FEATURES             Location/Qualifiers
     source          1..5850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1884..1955
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    1977..2565
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2030..2050)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2448..2459)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2597..3073
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2796..2815)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3113..3253
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3113..3242
                     /label=AAV2 ITR
     rep_origin      3328..3783
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3415..3434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3625..3646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3800..3819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3919..3941
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3979..3997)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4065..4169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4170..4238,4239..5030)
                     /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(4388..4407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5201..5789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5690..5709
                     /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 gcgttgggaa gtgtccaggg ccatggggtc acaactaaca
      181 gcccttggaa atgagcttgt gtccttaatc ttcatgacct aacgtgattt ctctagaaac
      241 atcagtgcat taacaggaag acaagatgga agatcatatt ttggctctcc ttccttggtg
      301 ggttgacact gctggtccta tccactagta aaagcatgac tcttaggctc tgtgtggcca
      361 gtggaaggtg gcagttggag gaagcagatg cttggccagc ctttgcctgg gagcagtcta
      421 gctctgagtg tcttattgga gcagctgctg gcatccagag ttctttttgg attcacgatt
      481 taattcaaaa gctttgtgct cccgagaatc agctctggtc tttcaaaaat aagatgtgag
      541 tccgccgaga ggctcccaag gtattgcctt gccaactgca agccttttag gagcagttta
      601 gtgagtggtg actgctagtt gcagttggct gttagcccag agctaataat agatagaaaa
      661 ggtatatact taaggagtct ggaaactgag gtttatctac tcacagaaaa tgagtttcta
      721 aaaaactagc ttgaaactta cccagaaaaa tcttagaaca tggttctcca atgtcaaggt
      781 aagtgttctg tgacactggg cttgaattat gtagggacca cagattttag aatttggacc
      841 cctgaacttg cttcacaccc caccaggaac cttcctgtac aacagccctc agaattcatc
      901 tacatggtct tttctcagta tgggatccgg tctagcaagt ggagcacacc ttctattgct
      961 taaagatttg tttatgtata tgggtatttt ggctgcatgc atatttgcac accaaaagaa
     1021 ggcagcggat cccatggaat tactgtgggt gctgggaatt gaactcagga cctctggaag
     1081 aatagccagt gctcttaacc actgagccat gcctgcagtc catctatttt ttattctagt
     1141 acagcccctc ttcattctta ctgaaatagt aatgcctgaa ccacacagct tcacatttag
     1201 ttacaaagaa agagtgggag tatcatgtga caattagtgt tgttgactct ccaggacaaa
     1261 cttatgggaa agggaggtct cctgacccct gaacaatcat tttacttgag gataattttc
     1321 attgcactca gaaacatgct gaattattgt ccttaccctt gccccatctc ttgctctggt
     1381 agagaatggc caaagcctgg aaacaggatg gcttgacaga agctctactt ggcttcccag
     1441 acccaagctg gattaaacca ggttccctaa ggagtcttaa ggcagctgcc aggagcaagg
     1501 ggcccactca ttggctattg gccttgactg ggttggccaa tggtaagctg gggtctgcct
     1561 gtccccatga gtaccagact aatgagacct ggccactttc tcctcatttc tgtctgtacg
     1621 attgtcagtg gatctgacga caccaaaagg taagaacaat tctatattct cggctggctg
     1681 ggtatgaatg cagaatccca cttgggctgt gttcagattt tgctctctgc agcagtgtga
     1741 aggtagatgt tgagaaaact caggccttgg aaacatggtt ggggcaaccg gtggtttagt
     1801 gaaccgtcag atccgctaga gatctgcggc cgcagtactg ctagcactag tgatatccga
     1861 tcggtcgacg cccgggcctc gaggactaca aggatgacga tgacaaggat tacaaagacg
     1921 acgatgataa ggactataag gatgatgacg acaaataaga tatcaagctt atcgataatc
     1981 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2041 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2101 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2161 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2221 ggggcattgc caccacctgt cagctccttt ccggaacttt cgctttcccc ctccctattg
     2281 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2341 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     2401 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2461 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2521 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     2581 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     2641 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     2701 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     2761 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     2821 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     2881 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     2941 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     3001 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     3061 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     3121 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     3181 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     3241 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3301 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     3361 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     3421 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3481 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3541 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3601 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     3661 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     3721 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3781 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3841 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3901 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3961 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4021 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4081 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4141 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4201 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4261 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4321 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4381 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4441 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4501 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4561 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4621 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4681 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4741 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4801 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4861 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4921 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4981 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5041 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5101 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5161 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5221 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5281 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5341 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5401 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5461 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5521 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5581 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5641 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5701 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5761 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5821 tggccttttg ctggcctttt gctcacatgt
//
