LOCUS       sequence-392388-        7258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted mouse CES2C S230A mutant .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7258)
  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 7258)
  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: pDEST40-CES2C-Delta-C-S230A
FEATURES             Location/Qualifiers
     source          1..7258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1211..1230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1402..1781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1782..1985
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1936..1956
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2030..2049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2030..2048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2078..2102
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2207..2230
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3833..3850
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3873..3897)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3923..3964
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3974..3991
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4014..4031)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4020..4244
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4290..4718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4377..4396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4587..4608
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4727..4747)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4732..5061
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4912..5047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4974..4993
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5128..5922
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5182..5201)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5792..5811
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6098..6219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6135..6154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6189..6208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6268..6284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6268..6284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6281..6303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6292..6308
                     /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)."
     promoter        complement(join(6316..6322,6323..6340,6341..6346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6361..6382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6499..6516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6670..7258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6750..6769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1261 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     1321 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     1381 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     1441 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1501 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1561 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     1621 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     1681 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     1741 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     1801 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     1861 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     1921 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     1981 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     2041 actataggga gacccaagct ggctagttaa gctatcaaca agtttgtaca aaaaagcagg
     2101 ctccgcggcc gcccccttca ccgccaccat gacacggaac caactacata actggctgaa
     2161 tgctgggttc ttcgggctgc tgcttcttct tatccatgtg cagggtgact acaaggatga
     2221 cgacgataag gggggcggtc aggactcacc agaggccaac cccatcagaa acacacatac
     2281 aggacagatc caaggcagcc ttatccacgt gaaggacact aaagctggtg tccacacctt
     2341 cctgggaatc ccctttgcca aacctcctgt aggaccactg cgatttgcac ctcctgaggc
     2401 ccctgaacca tggagtggtg tgagagatgg aactgcacat ccggccatgt gtctgcaaaa
     2461 tcttgacatg ctgaatgaag caggcctgcc agatatgaaa atgatgctgt cttccttccc
     2521 tatgtctgag gactgcctgt atctcaacat ctacacccca gcccatgccc atgagggctc
     2581 taacctgcct gtgatggtgt ggatccatgg aggtgcactg gttataggaa tggcttccat
     2641 gtttgatgga tctctattga cagtcaatga ggacttggtg gttgttacta tccagtatcg
     2701 tctgggtgtc ctgggctttt tcagcactgg agaccagcat gccagaggca actggggata
     2761 cctggaccaa gcagctgccc tacgctgggt ccagcagaac atcgctcact ttggaggcaa
     2821 ccctgaccgg gtcactattt ttggcgaggc tgcaggtggc acaagtgtgt cttcacatgt
     2881 tgtgtccccc atgtcccaag ggctcttcca tggtgccatc atggagagtg gggtggccct
     2941 gcttcctgac cttatctctg aaacctctga gatggtttcc actacggtgg ccaagctttc
     3001 tggatgtgag gctatggatt cccaggccct ggtgcgctgt ctgagaggca aaagtgaagc
     3061 agagattctg gctattaaca aggtcttcaa gatgatccct gctgtggtgg atggggagtt
     3121 cttccccagg catcccaaag agttgttggc atctgaggat tttcaccctg tccccagcat
     3181 cattggtgtc aacaatgatg agtttggctg gtccattcct gtggtcatgg gctctgctca
     3241 gatgataaaa ggaataacca gagagaacct gcaggctgtt ctgaaggata cagcagtaca
     3301 aatgatgctg cctcctgagt gtagtgacct gctaatggaa gagtacatgg gggacactga
     3361 ggatgcccag accctccaaa tacagttcac agagatgatg ggagatttca tgtttgtgat
     3421 ccctgcactc caagtagcac attttcaacg ctcccatgcc cctgtctact tctatgagtt
     3481 ccaacatcca cccagctact tcaaggatgt cagaccaccc cacgtgaagg ctgaccatgc
     3541 tgatgagatt ccttttgtct ttgcgtcctt cttctggggc atgaaacttg acttcactga
     3601 ggaggaggag ctgctgagca ggaggatgat gaagtactgg gccaactttg cacgacatgg
     3661 gaaccccaac agtgagggtc taccctactg gcctgtgatg gaccatgatg agcagtacct
     3721 gcaactggac atccagcctg ctgtgggtcg agccctgaag gctggaaggc tgcagttctg
     3781 gaccaagact ctgccccaga agatccagga gctaaaggct tctcaggaca agcatcatca
     3841 ccaccaccat tagtttaagg gtgggcgcgc cgacccagct ttcttgtaca aagtggttga
     3901 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     3961 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     4021 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     4081 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4141 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4201 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     4261 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4321 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4381 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4441 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4501 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4561 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4621 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4681 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4741 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4801 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4861 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4921 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4981 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     5041 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     5101 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5161 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5221 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5281 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5341 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5401 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5461 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5521 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5581 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5641 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5701 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5761 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5821 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5881 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5941 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6001 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6061 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6121 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6181 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6241 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6301 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6361 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6421 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6481 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6541 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6661 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6721 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6781 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6841 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6901 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6961 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7021 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7081 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7141 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7201 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaa
//
