LOCUS       sequence-392384-        6604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Secreted mouse CES2A S227A mutant driven by heptaocyte-specific 
            promoter in AAV viral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6604)
  AUTHORS   Wei W, Riley NM, Lyu X, Shen X, Guo J, Raun SH, Zhao M, Moya-Garzon 
            MD, Basu H, Sheng-Hwa Tung A, Li VL, Huang W, Wiggenhorn AL, 
            Svensson KJ, Snyder MP, Bertozzi CR, Long JZ
  TITLE     Organism-wide, cell-type-specific secretome mapping of exercise 
            training in mice.
  JOURNAL   Cell Metab. 2023 Apr 28:S1550-4131(23)00138-9. doi: 
            10.1016/j.cmet.2023.04.011.
  PUBMED    37141889
REFERENCE   2  (bases 1 to 6604)
  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: "Cell Metab.
            2023 Apr 28:S1550-4131(23)00138-9. doi: 10.1016/j.cmet.2023.04.011."
COMMENT     Sequence Label: pAAV-Tbg-CES2A-Delta-C-S227A
FEATURES             Location/Qualifiers
     source          1..6604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    49..70
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(85..90,91..108,109..115)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    123..139
                     /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     128..150
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     147..163
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     147..163
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   201..330
                     /label=AAV2 ITR
     polyA_signal    418..625
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     614..631
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             698..709
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1107..1127
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2789..2812)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2911..2930)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2913..3045
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(3105..3514)
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     repeat_region   3854..3983
                     /label=AAV2 ITR
     primer_bind     complement(4002..4019)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4002..4018)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4011..4033)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4160..4615
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4247..4266)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4457..4478
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4641..4745
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4746..4814,4815..5606)
                     /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(4964..4983)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5777..6365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6266..6285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
       61 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      121 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccagatt
      181 taattaaggc cttaattagg ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
      241 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      301 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact
      361 tatctacgta gccatgctct aggaagatcc ttatcgggaa ttcgccctta actcgagtcc
      421 ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
      481 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
      541 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
      601 gatggctggc aactagaagg cacagtcgag gcagatctcg aagacgcgga agaggccgca
      661 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
      721 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg
      781 atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
      841 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
      901 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
      961 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
     1021 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     1081 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     1141 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt ggatccaagc ttctacttgt
     1201 cctgagaacc cttgagctcc tggatcttct ggggcagagt cttggtccag aactgcagcc
     1261 ttctggcctt cagggctcga cccacagcag gctgggtgtc cagctgcagg tactgctcat
     1321 catggtccaa cacaggccag gagggtagac cctcactgtt ggggttccca tttcttgcga
     1381 aattggccca gtacttcatc atcatcctct ttagcagctc ctcctcctca gtgaggtcca
     1441 aattcatgtc ccacaaatag gagccaaaga caaaggcaac gtcatcacca tggtctgcct
     1501 tcacatgtga tggcctgaca tgcttgatga agctggacag atgctggaac tcatagaagt
     1561 agacaggagc ctgggaacgc tggaaatgtg ccacttggag agcagggatc acaaacatga
     1621 agtcccccag catctctctg aactgtgctt gcagggtctc tgggtcctca gtgtccccca
     1681 tatactcctc cactagcagg tgactacatt ctggaggcaa catcatccgt gctgctgtgt
     1741 tcttcagaac agctggcagg gtctctctgg ttatgttctt tatgatatgg tcaaggccca
     1801 tgaacatggg gactccccag ccacattcat ctgtgttgac accaatgatg ctggggacag
     1861 ggtgaaaatc catagaggtc agcagctcct gaggatgctt gggtaggaac tccccatcca
     1921 ccacagcagg gatcatcttg aagacctggt taatagccag gatctcttgt ttactctttg
     1981 ctctcaggca gtggatcagg gcttctgagt ctgtggcttc acaaccagat aggttggcta
     2041 ctgtcttgta gaccacctca gaagtgtcag agatgaggtc aggcagcagg gccactccac
     2101 tctgcataat ggctccatga aagagtcctt tggacattgg ggacagaata tgagaagaca
     2161 cacttgtacc tcctgctgcg actccaaata tagtgaccct gtcacggttg cctccaaagt
     2221 aagcaatatt cttctgcacc cagcgtaagg cagccacttg gtccaggtat ccccagttgc
     2281 ctctggcatg ctggtctcca gtgctgaaga agccaaggac ccccaagcga tactggatgg
     2341 cgacaattac tatttcctca gtggctgcca gtttagatac atcattcatg gaagctgatc
     2401 ctacaaccag tccaccaccg tggatccata ccatcacagg caggttagag ccctcctggg
     2461 cgtgggttgg tgtgtagatg ttgagataca ggcagtcctc ggacatagag atgggaggca
     2521 ggatcagatt catttccttc gcatcctcta aattcattat atcaggctgc agacacatgg
     2581 ccggctgtga ggtcccatct ctcacaccac tccatggctc agggtcctca ggaggtgcaa
     2641 agcgcagtag tcctacagga ggcttggcaa aggggattcc caggaaggca tggacaccag
     2701 atttagtgtc cttcacgtgg acaaagctgc ctctcacctg acccctgtgg gtgttcctga
     2761 tggggctggc tgagtcctga ccgcccccct tatcgtcgtc atccttgtag tcgccatgca
     2821 catgctggag gagaagcagg agcccacagg caacaacaca cagccaacca ggaagtctag
     2881 ccaatggcat ggtggcgcgg ccgcctggac acctgtggag agaaaggcaa agtggatgtc
     2941 agtaagacca ataggtgcct atcagaaacg caagagtctt ctctgtctcg acaagcccag
     3001 tttctattgg tctccttaaa cctgtcttgt aaccttgata cttacctgcc cagtgcctca
     3061 cgaccaactt ctgcagtctc tcagaaagca acatctttcc atttttatag catgtcctgt
     3121 attgcaaaac tagagcactc ttagttataa attaacccca gcattaaccc tgggatggta
     3181 ttttcaaagc tgatgtacaa aaccaaggcc aagaggaaca aagcaaaaaa tgataaggtt
     3241 tgaactttga gtgattgctt tggctgccag acccttcatg taaaacaaaa gagaaagagg
     3301 attgtcagag ctggaggggt ttaagtccat gctggcaaga gtgtcttcag caggcagaat
     3361 aggggccata ggcaaaagca ccaagaggca gcaggaaaaa gttctcacta ttgggccaaa
     3421 cagcagtgga attggcagtt ttttaaggga aagttgtaca aaagcagagg ctgggtgatc
     3481 ctttctaata ggttctgtag aaattataca tgcattcgca gaggaaatga tgtcaaaggt
     3541 agcttccagc cctggcgcgc cggatctgga atgtttgtgc tcctgagatt attaaccaga
     3601 gcaaacagag agagtaaatg ctgccaaggg ccacttggac ttttgactgc tttttaaaaa
     3661 ttaacctgga tctggaatgt ttgtgctcct gagattatta accagagcaa acagagagag
     3721 taaatgctgc caagggccac ttggactttt gactgctttt taaaaattaa cctgctagct
     3781 taagggcgaa ttccgatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3841 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3901 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3961 tcagtgagcg agcgagcgcg cagccttaat taacctaatt cactggccgt cgttttacaa
     4021 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4081 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4141 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4201 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4261 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4321 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4381 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4441 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4501 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4561 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     4621 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4681 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4741 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4801 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4861 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4921 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4981 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5041 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5101 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5161 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5221 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5281 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5341 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5401 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5461 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5521 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5581 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5641 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5701 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5761 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5821 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5881 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5941 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6001 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6061 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6421 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6481 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6541 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6601 gctg
//
