LOCUS       sequence-371300-        6674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lipophagy-inducing adaptor protein, AAV, negative control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6674)
  AUTHORS   Minami Y, Hoshino A, Higuchi Y, Hamaguchi M, Kaneko Y, Kirita Y, 
            Taminishi S, Nishiji T, Taruno A, Fukui M, Arany Z, Matoba S
  TITLE     Liver lipophagy ameliorates nonalcoholic steatohepatitis through 
            extracellular lipid secretion.
  JOURNAL   Nat Commun. 2023 Jul 13;14(1):4084. doi: 10.1038/s41467-023-39404-6.
  PUBMED    37443159
REFERENCE   2  (bases 1 to 6674)
  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: "2023-07-13"; volume: "14(1):4084. doi"; pages: " 
            10.1038/s41467-023-39404-6"
COMMENT     Sequence Label: pAAV.TBG mCherry-LDTS-LIRmut
FEATURES             Location/Qualifiers
     source          1..6674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        470..879
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     intron          939..1071
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1054..1073
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1083..1092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1089..1796
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1234..1252)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1502..1521
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2838..2864
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2927..2947)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3345..3356)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3423..3440)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3429..3636
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3724..3853
                     /label=AAV2 ITR
     primer_bind     complement(3872..3889)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3872..3888)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3881..3903)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4030..4485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4117..4136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4327..4348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4511..4615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4616..4684,4685..5476)
                     /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(4834..4853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5647..6235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6136..6155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6523..6544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6559..6564,6565..6582,6583..6589)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6597..6613
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6602..6624
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6621..6637
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6621..6637
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg gaattcgccc ttaagctagc aggttaattt ttaaaaagca gtcaaaagtc
      241 caagtggccc ttggcagcat ttactctctc tgtttgctct ggttaataat ctcaggagca
      301 caaacattcc agatccaggt taatttttaa aaagcagtca aaagtccaag tggcccttgg
      361 cagcatttac tctctctgtt tgctctggtt aataatctca ggagcacaaa cattccagat
      421 ccggcgcgcc agggctggaa gctacctttg acatcatttc ctctgcgaat gcatgtataa
      481 tttctacaga acctattaga aaggatcacc cagcctctgc ttttgtacaa ctttccctta
      541 aaaaactgcc aattccactg ctgtttggcc caatagtgag aactttttcc tgctgcctct
      601 tggtgctttt gcctatggcc cctattctgc ctgctgaaga cactcttgcc agcatggact
      661 taaacccctc cagctctgac aatcctcttt ctcttttgtt ttacatgaag ggtctggcag
      721 ccaaagcaat cactcaaagt tcaaacctta tcattttttg ctttgttcct cttggccttg
      781 gttttgtaca tcagctttga aaataccatc ccagggttaa tgctggggtt aatttataac
      841 taagagtgct ctagttttgc aatacaggac atgctataaa aatggaaaga tgttgctttc
      901 tgagagactg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
      961 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1021 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccagg
     1081 cggccgccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     1141 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     1201 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     1261 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     1321 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     1381 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     1441 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     1501 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     1561 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     1621 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     1681 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     1741 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggag
     1801 gtggaagcat cgagaaggtg gtagagttcc tcctgccacc agacaaggag tcagcccctt
     1861 cttccggacg gcagaggacc cagaaggctc ccaaggccaa accaagcctt gtgaggaggg
     1921 tcagcaccct ggccaacact ctttctcgac acaccatgca aaccacagca tgggccctga
     1981 agcagggcca ctctctggcc atgtggatcc cgggtgtggc acccctgagc agcctggccc
     2041 agtggggcgc atcggcagcc atgcaggtgg tgtcccggcg gcagagtgag gtgcgggtgc
     2101 cctggctgca caacctggca gcctctcagg atgagagcca tgacgaccag acagacacag
     2161 agggagagga gacagacgac gaggaggagg aagaagagtc cgaggctgag gagaacgtgc
     2221 tcagagaggt tacagccctg cccaacccga gaggcctcct gggtggtgtg gtacacaccg
     2281 tgcagaacac tctccggaac accatctccg cagtgacctg ggcacctgcg gctgtgctgg
     2341 gcacggtggg aaggatcctg cacctcacac cagcccaggc tgtctcctct accaaaggga
     2401 gggccatgtc cctatccgat gccctgaagg gtgttacgga taacgtggta gacactgtgg
     2461 tacactatgt gccgcttccc aggctgtccc tgatggagcc cgagagcgaa ttccgagaca
     2521 tcgataaccc ttcagcagag gcggagggat ccggtggagg cggaggttct gggggaggag
     2581 gtagtggcgg tggtggttca ggaggcggcg gaagcttgga tcctaggtca tctgaggacc
     2641 ccactgatga ctccaggctt cccagggccg aagcggagca ggaaaaggac cagctcagga
     2701 cccaggtggt gaggctacaa gcagagaagg cagacctgtt gggcatcgtg tctgaactgc
     2761 agctcaagct gaacgcagct ggcgctgcag aagatgcagc agttgaagct aggatggctg
     2821 aaggagaagc agaagggtac ccatacgatg ttccagatta cgcttgagga tccaatcaac
     2881 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2941 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3001 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3061 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3121 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3181 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3241 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     3301 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3361 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgagatc
     3421 tgcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3481 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3541 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3601 ggaggattgg gaagacaata gcaggcatgc tggggactcg agttaagggc gaattcccga
     3661 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     3721 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3781 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3841 agcgagcgcg cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact
     3901 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3961 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4021 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4081 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4141 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4201 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4261 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4321 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4381 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4441 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4501 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4561 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4621 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4681 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4741 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4801 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4861 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4921 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4981 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5041 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5101 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5161 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5221 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5281 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5341 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5401 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5461 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5521 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5581 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5641 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5701 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5761 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5821 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5881 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5941 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6001 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6061 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6121 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6181 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6241 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6301 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6361 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6421 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     6481 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     6541 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     6601 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca gatttaatta
     6661 aggccttaat tagg
//
