LOCUS       sequence-425915-        6096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human NWD1 fragment P859 to V1462. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  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 6096)
  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_NWD1:859-1462
FEATURES             Location/Qualifiers
     source          1..6096
                     /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    3159..3280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3170..3189)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3395..3428)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3438..3485
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3517..4311
                     /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(3571..3590)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4181..4200
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4469..5057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4549..4568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5183..5231
                     /note="synthetic polyadenylation signal"
     CDS             complement(5255..5653)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5650..5659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5684..6041)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5698..5833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5850..6041
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5998..6018
                     /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 tcgagcggtc ccctccgggc
     1261 aactctcagc ggctgtcaca aaggcatcac cgccatggca tggggtgtgg aggagaagct
     1321 gctggtgatt ggcacccagg atggcatcat ggctgtgtgg gacatggaag agcagcatgt
     1381 gatccacatg ctaactggac acacaggaga ggtgaggtgt gtgaaaatat ttgccaaagg
     1441 gaccctcgcc atctctgctt caaaggatta cacgctgcac ttgtggaact tactctctgg
     1501 ccaggagaaa tttaccattt gggatggagg ctcaaaaaat cccgctgaac ctcagatctg
     1561 gaaccttcat gtggatgagg cacacaaagt tgtgtattca gcatctggct caaagatcaa
     1621 tgcttggaat ctggaaactg cagagccggt attccatatc ctgggagatg cctctgatcc
     1681 ttggatgtgc atggccgtgc tggcctccca ggccacactg ctgacagtgt ccagggatgg
     1741 tgtggtcagt ctgtggagct cagctacggg aaaacttcag gggaagcaac atatgtccag
     1801 catcaaagaa gaaacaccta cctgtgccgt ctcagtccag aagcaaggaa agcttgttac
     1861 cgggtttagc aatggctcca tctctttggt ttcctccaaa ggggacagat tgctggagaa
     1921 gcttccagat gctgtgaggt tcctggtggt ctctgaagat gagtccctcc tcgccgcagg
     1981 ctttggaaga tcggtgcgga tattcttggc ggactcgagg ggctttcgcc gattcatggc
     2041 catggatctg gaacatgaag acatggtgga gacggctgtt tttggtactg agaacaacct
     2101 gatcatcacg gggtcccttg atgcgctcat tcaggtgtgg agtctgtcag aacaggggac
     2161 ccttctggac atcctggaag gcgtcggggc ccccgtgagc ctgctggccc gcggcggggc
     2221 tttggtggca tctgcttccc cacagtcctc atctttcaag gtctgggatc tcagcgatgc
     2281 tcataggtcc cgggtgcctg caccatttct ggaccgcacc ggcctcaccg cagtgtccca
     2341 caatggaagc tacgtctact tccccaaaat tggggacaaa aacaaagtca ctatttggga
     2401 cttggcagaa ggcgaggaac aagattccct ggacacctcc agtgagatca ggtgtctgga
     2461 ggttgctgag cagcgcaagc tcctatttac gggcctcgtg tcgggggtcg tccttgtgtt
     2521 ccccctgaat tccaggcagg acgtgatatg cattccccct cccgaggccc ggaaagcaat
     2581 caactgcatg tccctgagca agtgcgagga ccgcctggcc atcgcctatg acaacatcgt
     2641 cctggtgctg gacatcacct ccggggaccc ctgcccggtc atcgatgggc caagatacac
     2701 cttttacact cagctgcccg agaccctctc cagcgtggcc attctgacgg actaccgcgt
     2761 ggtctacagc atgaccaatg gggacctctt tctttacgag tgtgcaactt ccaaagcgtt
     2821 tcccttggag acccacagga gccgagttgc ctgtgtggag gtcagccaca aggagcagct
     2881 ggtggtcagc gggtctgagg atgccctgct gtgtctctgg gacctgcagg cacgcaagtg
     2941 gaaattcgag atgagctaca cgagttctta ctgcagaggg gtccaatgtg cttgcttctc
     3001 caaggatgac aagtatgtgt acgtgggctt aaaggatcgc tctatacttg tttggagtgt
     3061 gtaatctaga ggtcaagctt cgggggttta aacctgcagg catgcaagct gatttggctg
     3121 ctgccaccgc tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3181 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3241 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3301 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3361 caaatgtggt aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac
     3421 gaagttatgg ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat
     3481 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     3541 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3601 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3661 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3721 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3781 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3841 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3901 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3961 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4021 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4081 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4141 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4201 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4261 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg
     4321 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     4381 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4441 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4501 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4561 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4621 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4681 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4741 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4801 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4861 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4921 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4981 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5041 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5101 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5161 tccttttata gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     5221 gatattttat tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg
     5281 gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt
     5341 tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc
     5401 ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag
     5461 cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac
     5521 cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga
     5581 tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca
     5641 aaggcttggc catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc
     5701 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     5761 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     5821 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     5881 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     5941 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     6001 ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc
     6061 ctctttcaga ggttatttca ggccatggtg ctgcgc
//
