LOCUS       sequence-423398-        6441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human NWD2 fragment S944 to S1662. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6441)
  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 6441)
  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_NWD2:944-1662
FEATURES             Location/Qualifiers
     source          1..6441
                     /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"
     regulatory      2162..2171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3504..3625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3515..3534)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3740..3773)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3783..3830
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3862..4656
                     /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(3916..3935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4526..4545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4814..5402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4894..4913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5528..5576
                     /note="synthetic polyadenylation signal"
     CDS             complement(5600..5998)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5995..6004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6029..6386)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6043..6178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6195..6386
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6343..6363
                     /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 tcgagcggtt ccattgtacc
     1261 actgcattca tccatggatg tgacatacag cccagagcgt cttcccttat catccagtca
     1321 cctgcatgtc acagagatcc tgcctacctg taaccccagc actgtcctca cagctttaga
     1381 aaatggttcc atcagcacct gggatgtaga gactcgacag ctactcaggc aaatcaccac
     1441 agcccagtct gtcatcctgg gcatgaaact caccagtgat gaaaagtacc ttgtggtggc
     1501 tacaacaaat aacaccttgt tgatttatga caatgtcaat tcttgcctcc tgtctgaagt
     1561 agaaatcaag gggaccaagc atggaagcag cgccacctac atcaatggat ttacactgtc
     1621 cgccaaccac gcccttgcat ggctcgaagc cagcaaagat gtcactgtca tcgatctgct
     1681 gtacggatgg ccgctttacc agttccactg ctggtatgaa gtgacgtgcg tccagtgctc
     1741 cctggatggt ctgtatgctt tctgtggcca atacctgaac acaaccacca tatttcattt
     1801 agggagtgga gaaaagttat gtacagtgac atcagaattt tcaggtggat ttgtgaagtt
     1861 tcttcttatc ttggacacag ctcaggaaat ggtcatggta gacagtgaag gaagtctttc
     1921 tgtttggaat actgaggaca tttccagccc ccagctgact gatgactttg attgccgaag
     1981 agaagacagt gaggtggtca gcattgagct ttcagaagac caaagtgcag ttctgatctg
     2041 taaagccctc agcattgagc tcttagatac tggtctgtgg aaggtggctg aaaagttcag
     2101 agccaagcac aacgaacgct ttatatctgc cgtgctgtct aaaaatggag attgtatcat
     2161 cgccaccatg gaaaatacct cagctgtgtt tttttggagg cgggacacag gacagtgtat
     2221 ggcaagcttg caggaaatct caggttccat tgttaagtta gtgaaatcca gtcaccacaa
     2281 tatgctactg tctttatcaa ccagtggtgt tctttccatt tgggacatag atataatcac
     2341 agctatgtcc aacatagata agactggaaa acccatccaa agtctgttgt tgcctgctag
     2401 aggggaaatc atttactccc tggatggatc cgattgtgtt cataagtgga acttcagcag
     2461 tggcttcatc gaagcagtat tcaagcatga aggaatagtt gaacactgtg tgttaacatc
     2521 caccggagat ataatggtga catcagatga caaaagcagc cagtatgtct ggcacaccag
     2581 cagtggtgaa aacctttttc gaattaacgg gcagagaata tctcagctgc tgattacaca
     2641 caatgaccag tttgtggtct cgctctgtga ggaaaatgcc tccagggttt ggaggctcgc
     2701 cacaggccac agggtctgca acattctgac cactttgcag aatgccttta ttacctccgc
     2761 aaataccttc gtggtgggaa tgactaaaag caaagtgttg gcagtcagtc tttggacagg
     2821 aagtatcacc aagaaatttt gctgtgaaga tgggaccacc atcgtgaatt ttaaattaat
     2881 ccctgactgt cctgatatca tcgtgtttat cacatcggcc gagactgtga acatctggag
     2941 tctgacagat gaagtgatct gtcggcgcgt gcaacttcca aacaacttct tgaaaaatct
     3001 ggaggacttt gaaatttctc ccaatggaaa gctaggcatt atagccaggg gagatgaaaa
     3061 catcaatgtg ctagatttat acagtggtaa attgcgggtg gttcacgcct ctgggatcat
     3121 ctggcggcag aggttgtctc gggatggtcg ctacctggta tacatttgtt tccgaaatgg
     3181 ggaggaggag gatgaaaatg gtgcaatatt cagtttaatt gtaatgagac tggcagatgg
     3241 caaaaatatc ggtgcttgtt ccctttacaa aacaccaact ttccttgcac tctcccagag
     3301 gcacctgaac atcattgtgg gctttgatga tgggagtata gggatctaca cggtagtaga
     3361 ccgtgtagat gctgcactga aaatcaaaat tgccacttca aatagctaat ctagaggtca
     3421 agcttcgggg gtttaaacct gcaggcatgc aagctgattt ggctgctgcc accgctgagc
     3481 aaggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3541 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3601 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3661 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3721 cttaattaag acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca
     3781 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3841 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4021 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4381 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4621 atcgccttct atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg
     4681 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5221 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     5401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     5521 gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     5581 ccatcgtgat ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa
     5641 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
     5701 gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc
     5761 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
     5821 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
     5881 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
     5941 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg
     6001 tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc
     6061 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     6121 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     6181 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     6241 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     6301 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     6361 tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta
     6421 tttcaggcca tggtgctgcg c
//
