LOCUS       sequence-347620-        5633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP under the control of distal enhancer and the promoter 
            of human adiponectin gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5633)
  AUTHORS   Shamsi F, Xue R, Huang TL, Lundh M, Liu Y, Leiria LO, Lynes MD, 
            Kempf E, Wang CH, Sugimoto S, Nigro P, Landgraf K, Schulz T, Li Y, 
            Emanuelli B, Kothakota S, Williams LT, Jessen N, Pedersen SB, 
            Bottcher Y, Bluher M, Korner A, Goodyear LJ, Mohammadi M, Kahn CR, 
            Tseng YH
  TITLE     FGF6 and FGF9 regulate UCP1 expression independent of brown 
            adipogenesis.
  JOURNAL   Nat Commun. 2020 Mar 17;11(1):1421. doi: 10.1038/s41467-020-15055-9.
  PUBMED    32184391
REFERENCE   2  (bases 1 to 5633)
  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: "Nat 
            Commun."; date: "2020-03-17"; volume: "11(1):1421. doi"; pages: " 
            10.1038/s41467-020-15055-9"
COMMENT     Sequence Label: pAAV-hAdipoq-GFP
FEATURES             Location/Qualifiers
     source          1..5633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(890..892,893..895,896..1609)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(890..892,893..895,896..1606)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(935..956)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1196..1215)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1543..1564
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1628..2216
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1681..1701)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2099..2110)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2230..2364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2254..2273)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2308..2327
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2450..2467)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2456..2663
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2672..2801
                     /label=AAV2 ITR
     rep_origin      2887..3342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2974..2993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3184..3205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3409..3428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3528..3550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3588..3606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3674..3778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3779..3847,3848..4639)
                     /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(3997..4016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4810..5398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5299..5318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggctgcagag agggagtggc caactccatc actaggggtt cctcagatct gaattcggta
       61 cctctttcca catgacggcc tttgtggtgg gtggcagatt gccctgaggc ctcgcaaaat
      121 gctaggcttt cacaatgtca ctgactgaca gccaggccca gcacagtctt ggtgtgattg
      181 tggggctaaa gttattccac cttgtgcaat agctacagcg aattcatgtg caaggtcttt
      241 cctaaatcca gggtccaggt cagagagtgg aggatgtgct ctatttctta cctgattgca
      301 gacccctctg acagtgctcc cttctgaagc actcactgtc tgaacgtaca cagtctcaga
      361 cttaatcatg cacagtgagc aagactgtgg tgtgataatt ggcgtccctg acttattagg
      421 gcaaatctat gggaggggga gacctcctgg accactgagc aattaattca tttacattag
      481 gaagtttctc cgtcagatgc aggaaaaaaa tcttgttttc ctgctgtggt tttgactttt
      541 gccccatctt ctgttgctgt tgtaggaggc aaaataaggg tcaaggcctg gaaacacaag
      601 tgctttgact gaagctccac ttggcttccg aagcccaagc tgggttgtac caggttccct
      661 agggtgcagg ctgtgggcaa ctgccaggga catgtgcctg cccaccggcc tctggccctc
      721 actgagttgg ccaatgggaa atgacaattg tgaggtgggg actgcctgcc cccgtgagta
      781 ccgggctgtt gaggctgggc catctcctcc tcacttccat tctgactgca gtctgtggtt
      841 ctgattccat accagagggt aaaaactagt ccgcggaatt ctcgagacca tggtgagcaa
      901 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      961 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1021 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1081 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1141 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1201 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1261 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1321 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1381 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1441 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1501 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1561 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag cggccgctct
     1621 agaagataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1681 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1741 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     1801 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     1861 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     1921 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     1981 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     2041 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2101 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2161 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     2221 gactagagag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2281 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2341 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2401 cagggggagg tgtgggaggt tttttagtcg actagagctc gctgatcagc ctcgactgtg
     2461 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2521 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2581 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2641 gacaatagca ggcatgctgg ggagagatct gaggaacccc tagtgatgga gttggccact
     2701 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2761 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gagagggagc tgcagcctgg
     2821 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2881 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2941 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3001 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3061 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3121 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3181 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3241 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3301 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     3361 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     3421 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3481 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3541 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3601 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3961 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4021 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4081 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4321 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4381 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4621 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5461 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5521 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5581 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gca
//
