LOCUS       sequence-426148-        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect expression of human PPWD1 fragment A80 to K453. N-terminal 6X
            His tag with TEV cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 5855)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pFBOH-MHL_PPWD1:80-453
FEATURES             Location/Qualifiers
     source          1..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(255..788)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(958..963,964..980,981..986))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        1338..1429
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1354..1371
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1488..1505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1518..1538
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2798..2932
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2835..2854)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2889..2908
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2961..3126
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3310..3764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3397..3416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3606..3627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3790..3894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3895..3963,3964..4755)
                     /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(4113..4132)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4926..5514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5415..5434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5668..5685
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  join(5819..5855,1..188)
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
ORIGIN
        1 gatctaaact atgacaataa agtcttaaac tagacagaat agttgtaaac tgaaatcagt
       61 ccagttatgc tgtgaaaaag catactggac ttttgttatg gctaaagcaa actcttcatt
      121 ttctgaagtg caaattgccc gtcgtattaa agaggggcgt ggccaagggc atggtaaaga
      181 ctatattcgc ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg
      241 gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc
      301 cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg
      361 tagatcacat aagcaccaag cgcgttggcc tcatgcttga gcagattgat gagcgcggtg
      421 gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata gatctcacta
      481 cgcggctgct caaacttggg cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg
      541 tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag gtaatcggag
      601 tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa aagatcaaga
      661 gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat gatgcccata
      721 cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctc
      781 cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata
      841 gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg
      901 ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagt
      961 ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct tcatccgttt ccacggtgtg
     1021 cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg gctggcgaac
     1081 gagcgcaagg tttcggtctc cacgcatcgt caggcattgg cggccttgct gttcttctac
     1141 ggcaaggtgc tgtgcacgga tctgccctgg cttcaggaga tcggaagacc tcggccgtcg
     1201 cggcgcttgc cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa
     1261 ggcgagcatc gtttgttcgc ccaggactct agctatagtt ctagtggttg gctacgtata
     1321 ctccggaata ttaatagatc atggagataa ttaaaatgat aaccatctcg caaataaata
     1381 agtattttac tgttttcgta acagttttgt aataaaaaaa cctataaata ttccggatta
     1441 ttcataccgt cccaccatcg ggcgcggatc tcggtccgaa aaccatgcat catcatcatc
     1501 atcacagcag cggcagagaa aacttgtatt tccagggcgc atccatgtat gagcgcagtt
     1561 acatgcatag agatgttatc acccatgtgg tatgcaccaa aacagatttt attattactg
     1621 ccagtcatga tggacatgtc aagttctgga aaaaaataga agagggaatt gaatttgtta
     1681 aacattttcg tagtcacctg ggagttattg agagtattgc agttagctct gagggagcat
     1741 tgttctgttc tgtgggtgat gataaagcaa tgaaggtgtt tgatgtagtg aactttgaca
     1801 tgatcaacat gctgaaactt ggctattttc ctggacagtg tgagtggatc tattgcccag
     1861 gggatgcaat ttcttcagtt gctgcttccg aaaagagtac aggaaaaatt ttcatttatg
     1921 atggccgagg agataaccag ccacttcata tttttgacaa actccataca tcacctctta
     1981 ctcagatacg gctaaaccca gtttacaaag cagtagtgtc ttctgacaaa tctgggatga
     2041 ttgaatactg gactgggcct cctcatgaat ataaattccc caaaaatgtg aactgggaat
     2101 ataaaactga cactgattta tatgaatttg ccaagtgtaa ggcttatcca accagcgtat
     2161 gtttttcacc agatgggaag aaaatagcta ctattggttc tgatagaaaa gttagaattt
     2221 tcagatttgt aactggaaaa ctcatgagag tctttgatga atcactaagc atgtttactg
     2281 aactgcaaca gatgaggcaa cagttaccag acatggaatt tggccgacga atggctgtag
     2341 aacgtgagtt ggagaaggtt gatgctgtaa gattaattaa tatagttttt gatgaaactg
     2401 gacacttcgt gctgtatgga acaatgctgg gcattaaagt tataaatgta gagacaaacc
     2461 ggtgtgtgcg gattttaggc aaacaagaaa atattagagt gatgcaattg gctttgttcc
     2521 aggggatagc caaaaagcat cgtgctgcaa ctactataga aatgaaagct tctgaaaatc
     2581 ctgttcttca gaatattcaa gctgacccaa caatagtctg tacatcattc aaaaagaata
     2641 gattttatat gtttaccaaa tgatgacgaa gcttgtcgag aagtactaga ggatcataat
     2701 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2761 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2821 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2881 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact
     2941 gatatcgcct aggagatccg aaccagataa gtgaaatcta gttccaaact attttgtcat
     3001 ttttaatttt cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt
     3061 ccctaaataa tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg
     3121 cccacagcgg ggcatttttc ttcctgttat gtttttaatc aaacatcctg ccaactccat
     3181 gtgacaaacc gtcatcttcg gctacttttt ctctgtcaca gaatgaaaat ttttctgtca
     3241 tctcttcgtt attaatgttt gtaattgact gaatatcaac gcttatttgc agcctgaatg
     3301 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3361 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3421 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3481 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3541 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3601 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3661 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3721 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     3781 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3841 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3901 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3961 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4021 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4081 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4141 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4201 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4261 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4321 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4381 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4441 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4501 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4561 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4621 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4681 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4741 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4801 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4861 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4921 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4981 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5041 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5101 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5161 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5221 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5281 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5341 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5401 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5461 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5521 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5581 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5641 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5701 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat agaccagccg
     5761 cgtaacctgg caaaatcggt tacggttgag taataaatgg atgccctgcg taagcgggtg
     5821 tgggcggaca ataaagtctt aaactgaaca aaata
//
