LOCUS       sequence-459311-        5916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cre recombinase from a rat Ucp1 enhancer-promoter. 
            Contains miR122 target sequences that suppress expression in 
            hepatocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5916)
  AUTHORS   Behrens J, Braren I, Jaeckstein MY, Lilie L, Heine M, Sass F, Sommer
            J, Silbert-Wagner D, Fuh MM, Worthmann A, Straub L, Moustafa T, 
            Heeren J, Scheja L
  TITLE     An efficient AAV vector system of Rec2 serotype for intravenous 
            injection to study metabolism in brown adipocytes in vivo.
  JOURNAL   Mol Metab. 2024 Oct;88:101999. doi: 10.1016/j.molmet.2024.101999. 
            Epub 2024 Jul 31.
  PUBMED    39094948
REFERENCE   2  (bases 1 to 5916)
  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: "Mol 
            Metab."; date: "2024-10"; volume: "88:101999. doi"; pages: " 
            10.1016/j.molmet.2024.101999. Epub 2024 Jul 31"
COMMENT     Sequence Label: pAAV-Ucp1-Cre-miR122
FEATURES             Location/Qualifiers
     source          1..5916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..82)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      876..1464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1365..1384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1526..1655
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             2410..2430
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3568..4156
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3621..3641)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4039..4050)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4188..4664
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4387..4406)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4704..4844
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4704..4833
                     /label=AAV2 ITR
     rep_origin      4919..5374
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5006..5025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5216..5237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5391..5410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5510..5532
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5570..5588)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5656..5760
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5761..5829,5830..5916,1..705)
                     /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"
ORIGIN
        1 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
       61 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
      121 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
      181 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
      241 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
      301 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
      361 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
      421 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
      481 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
      541 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
      601 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
      661 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
      721 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
      781 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
      841 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
      901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      961 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1021 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1081 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1141 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1201 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1261 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1321 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1381 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1441 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1501 ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag
     1561 gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     1621 agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgtg acgtcacagt
     1681 gggtcagtca cccttgatca cactgcacca gtcttcacct ttccacgctt cctgccagag
     1741 catgaatcag gctctctggg gataccggcc tcacccctac tgaggcaaac tttctcccac
     1801 ttctcagagg ctctgagggc agcaaggtca gccctttctt tggaatctag aaccactccc
     1861 tgtcttgagc tgacatcaca gggcaggcag atgcagcagg gaagggcctg ggactgggac
     1921 gttcatccta caagaaagct gtggaacttt tcagcaacat ctcagaaatc agatcgcact
     1981 tattcaaagg agccaggccc tgctctgcgc cctggtggag gctcctcatg tgaagagtga
     2041 caaaaggcac catgttgtgg atacggggcg aagcccctcc ggtgtgtcct ccaggcatca
     2101 tcaggaacta gtgccaaagc agaggtgctg gccagggctt tgggagtgac gcgcgtctgg
     2161 gaggcttgtg cgcccagggc acgcccctgc cgattcccac tagcaggtct tgggggacct
     2221 gggccggctc tgcccctcct ccagcaatcg ggctataaag ctcttccaag tcagggcgca
     2281 gaagtgccgg gcgatccggg cttaaagagc gagaggaagg gacgctcacc tttgagctcc
     2341 tccacaaata gccctggtgg ctgccacaga agttcgaagt tgagagttcg gggtaccgga
     2401 tccaccatgc ccaagaagaa gaggaaggtg tccaatctcc tgactgttca ccagaacctc
     2461 cctgcgctgc cagtagatgc cactagcgat gaggtcagga aaaatctcat ggatatgttt
     2521 agggatagac aggcgttttc tgaacacacc tggaaaatgc tgcttagcgt gtgccgatcc
     2581 tgggcagcct ggtgtaagct gaacaatcgc aaatggttcc ccgccgagcc ggaggacgtg
     2641 cgcgattacc tgctgtatct ccaggcaaga gggctggctg tcaagactat ccagcagcac
     2701 ttgggccaac tgaatatgct gcatcgacgc agcgggctcc cccggcctag cgattcaaac
     2761 gcagtctccc ttgttatgag gagaattaga aaggaaaacg tagatgcggg tgagagggct
     2821 aagcaggctc tcgcttttga gcggactgat ttcgaccagg tcagatccct gatggagaac
     2881 agcgatcggt gccaggacat caggaacctc gcatttctgg gaattgcata taacacactt
     2941 ctgcgcatag ctgagatcgc ccggatcaga gtgaaagaca tcagtcgaac ggacggcggc
     3001 cggatgctta ttcatattgg acgcacaaag acattggtca gcaccgctgg cgttgaaaag
     3061 gccttgtccc tgggcgtaac gaagctggtg gaaagatgga tctcagtgtc cggcgtggct
     3121 gacgacccta ataattactt gttctgtcga gtgagaaaaa acggagtcgc cgcgccctct
     3181 gccaccagcc aattgagtac acgggccctt gaagggatct ttgaggcaac ccaccgactc
     3241 atatacggag ccaaggatga cagtggccag aggtatctcg cctggtcagg tcattctgct
     3301 agggtggggg ccgcacgaga catggcgcgg gcaggagtct ccataccaga gattatgcaa
     3361 gctggaggtt ggacaaatgt gaacatcgtt atgaactata tccgcaatct tgactctgaa
     3421 accggggcca tggtgagact gctcgaagat ggtgactgaa ttcaccggtc aaacaccatt
     3481 gtcacactcc atactatcaa acaccattgt cacactccag atcaacaaac accattgtca
     3541 cactccacgc cggcgaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     3601 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3661 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3721 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3781 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3841 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3901 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3961 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
     4021 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4081 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4141 tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct
     4201 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     4261 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     4321 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     4381 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     4441 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     4501 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     4561 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     4621 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     4681 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     4741 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4801 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     4861 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     4921 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4981 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5041 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5101 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     5161 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5221 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     5281 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5341 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     5401 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5461 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5521 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     5581 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5641 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5701 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5761 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5821 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5881 cgagtgggtt acatcgaact ggatctcaac agcggt
//
