LOCUS       sequence-365645-        5624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the V16D mutant of human HSD17B11 with a C-terminal 
            FLAG-GFP tag. Confers G418 resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5624)
  AUTHORS   Demange P, Joly E, Marcoux J, Zanon PRA, Listunov D, Rulliere P, 
            Barthes C, Noirot C, Izquierdo JB, Rozie A, Pradines K, Hee R, de 
            Brito MV, Marcellin M, Serre RF, Bouchez O, Burlet-Schiltz O, 
            Oliveira MCF, Ballereau S, Bernardes-Genisson V, Maraval V, Calsou 
            P, Hacker SM, Genisson Y, Chauvin R, Britton S
  TITLE     SDR enzymes oxidize specific lipidic alkynylcarbinols into cytotoxic
            protein-reactive species.
  JOURNAL   Elife. 2022 May 10;11. pii: 73913. doi: 10.7554/eLife.73913.
  PUBMED    35535493
REFERENCE   2  (bases 1 to 5624)
  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: "Elife."; 
            date: "2022-05-10"; volume: "11. pii: 73913. doi"; pages: " 
            10.7554/eLife.73913"
COMMENT     Sequence Label: pEGFP-N1-HSD17B11-V16D-FLAGC
FEATURES             Location/Qualifiers
     source          1..5624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             967..990
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1000..1009
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1006..1008,1009..1011,1012..1725)
                     /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(1006..1008,1009..1011,1012..1722)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1051..1072)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1312..1331)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1659..1680
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1848..1969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1885..1904)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1939..1958
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1976..2431)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2113..2134)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2325..2344
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2458..2562
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2564..2921
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2564..2755
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2587..2607)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2772..2907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2956..3750
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2956..3750
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3010..3029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3620..3639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3938..3957)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3982..4029
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4359..4947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4848..4867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5122..5425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5426..5624,1..5)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5580..5600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagccacc
       61 atgaaatttc ttctggacat cctcctgctt ctcccgttac tgatcgactg ctccctagag
      121 tccttcgtga agctttttat tcctaagagg agaaaatcag tcaccggcga aatcgtgctg
      181 attacaggag ctgggcatgg aattgggaga ctgactgcct atgaatttgc taaacttaaa
      241 agcaagctgg ttctctggga tataaataag catggactgg aggaaacagc tgccaaatgc
      301 aagggactgg gtgccaaggt tcataccttt gtggtagact gcagcaaccg agaagatatt
      361 tacagctctg caaagaaggt gaaggcagaa attggagatg ttagtatttt agtaaataat
      421 gctggtgtag tctatacatc agatttgttt gctacacaag atcctcagat tgaaaagact
      481 tttgaagtta atgtacttgc acatttctgg actacaaagg catttcttcc tgcaatgacg
      541 aagaataacc atggccatat tgtcactgtg gcttcggcag ctggacatgt ttcggtcccc
      601 ttcttactgg cttactgttc aagcaagttt gctgctgttg gatttcataa aactttgaca
      661 gatgaactgg ctgccttaca aataactgga gtcaaaacaa catgtctgtg tcctaatttc
      721 gtaaacactg gcttcatcaa aaatccaagt acaagtttgg gacccactct ggaacctgag
      781 gaagtggtaa acaggctgat gcatgggatt ctgactgagc agaagatgat ttttattcca
      841 tcttctatag cttttttaac aacattggaa aggatccttc ctgagcgttt cctggcagtt
      901 ttaaaacgaa aaatcagtgt taagtttgat gcagttattg gatataaaat gaaagcgcaa
      961 acgcgtgatt acaaggatga cgacgataag ccaccggtcg ccaccatggt gagcaagggc
     1021 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1081 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1141 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1201 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1261 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1321 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1381 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1441 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1501 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1561 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1621 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1681 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     1741 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1801 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1861 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1921 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     1981 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2041 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2101 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2161 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2221 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2281 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2341 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2401 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2461 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2521 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2581 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2641 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2701 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2761 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2821 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2881 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     2941 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3001 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3061 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3121 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3181 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3241 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3301 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3361 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3421 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3481 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3541 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3601 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3661 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3721 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3781 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3841 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3901 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     3961 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4021 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4081 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac ccccccaagt
     4141 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4201 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4261 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4321 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4381 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4441 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4501 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4561 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4621 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4681 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4741 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4801 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4861 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4921 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4981 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5041 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     5101 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5161 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5221 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5281 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     5341 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     5401 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     5461 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     5521 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     5581 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agca
//
