LOCUS       sequence-425931-        5529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DAW1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5529)
  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 5529)
  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: pBiT3.1-N_DAW1:1-415
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2592..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2603..2622)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2828..2861)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2871..2918
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2950..3744
                     /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(3004..3023)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3614..3633
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3902..4490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3982..4001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4616..4664
                     /note="synthetic polyadenylation signal"
     CDS             complement(4688..5086)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5083..5092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5117..5474)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5131..5266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5283..5474
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5431..5451
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tgaagctcaa
     1261 gagcctcctg ctccggtatt acccgccagg aattatgttg gaatatgaaa aacatggaga
     1321 attaaagact aagtccatag atttgcttga tcttggtccc agcactgatg tcagtgcgtt
     1381 agtagaagaa atccagaagg cagaacctct actcacagct tcacgaacag agcaagtcaa
     1441 acttttgata cagaggttgc aagagaaact cggccagaac agcaatcaca cgttctatct
     1501 ttttaaggtt ctcaaagcac atatattgcc actgactaat gttgcactta acaaatcggg
     1561 ctcatgcttt atcacaggaa gctatgatcg gacgtgcaag ctctgggaca ctgcgtctgg
     1621 agaggagctg aacacgctgg agggccacag gaatgtggtt tatgccatag cattcaacaa
     1681 tccttacggt gacaaaatcg ccactgggtc ctttgataaa acttgtaaac tctggagtgt
     1741 ggaaacagga aaatgttacc ataccttcag gggtcataca gcagaaatag tgtgtttatc
     1801 atttaaccct caaagcacat tggtggcgac tggaagtatg gacacaacag ccaaattgtg
     1861 ggacattcag aatggcgagg aagtttacac cttaagagga cattctgccg aaatcatctc
     1921 cttgtcattt aacacctcag gagacagaat catcacgggg tcttttgatc ataccgttgt
     1981 agtgtgggac gctgatactg gaaggaaggt aaatatctta attggtcatt gtgctgagat
     2041 tagcagtgcc tcattcaatt gggattgctc tctaatatta actggctcta tggacaaaac
     2101 ctgcaagctg tgggatgcta caaatggaaa atgtgtggca accttaacag gccatgatga
     2161 tgaaatacta gacagctgct ttgattacac tggaaagctt attgcaactg cttcagctga
     2221 tggaacagca agaattttca gtgctgccac aagaaaatgc attgccaaac tggaaggtca
     2281 tgaaggtgaa atttcaaaga tttctttcaa ccctcaaggg aaccatcttc taactggcag
     2341 ctctgacaaa acggctagaa tctgggatgc tcagactggc cagtgcctcc aggttcttga
     2401 ggggcacact gatgaaatct tttcatgtgc tttcaactat aaaggcaaca tagtcattac
     2461 aggcagcaag gataatacct gtaggatatg gcgttaatct agaggtcaag cttcgggggt
     2521 ttaaacctgc aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg
     2581 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2641 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2701 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2761 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac
     2821 ttctaatata acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt
     2881 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     2941 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3001 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3061 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3121 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3181 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3241 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3301 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3361 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3421 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3481 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3541 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3601 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3661 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3721 cgccttcttg acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat
     3781 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3841 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3901 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3961 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4021 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4081 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4141 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4201 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4261 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4321 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4381 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4441 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt
     4501 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4561 tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac
     4621 aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg
     4681 ctagagttta gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg
     4741 tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca
     4801 cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga
     4861 tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac
     4921 aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga
     4981 gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg
     5041 ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt
     5101 cagaagaatc gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     5161 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     5221 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     5281 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     5341 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     5401 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt
     5461 tctttccgcc tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg
     5521 gtgctgcgc
//
