LOCUS       sequence-423465-        6488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human NWD2 fragment S944 to S1662. 
            C-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  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 6488)
  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-C_NWD2:944-1662
FEATURES             Location/Qualifiers
     source          1..6488
                     /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      2158..2167
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3551..3672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3562..3581)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3787..3820)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3830..3877
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3909..4703
                     /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(3963..3982)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4573..4592
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4861..5449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4941..4960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5575..5623
                     /note="synthetic polyadenylation signal"
     CDS             complement(5647..6045)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6042..6051
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6076..6433)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6090..6225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6242..6433
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6390..6410
                     /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 ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatgtccat tgtaccactg
     1261 cattcatcca tggatgtgac atacagccca gagcgtcttc ccttatcatc cagtcacctg
     1321 catgtcacag agatcctgcc tacctgtaac cccagcactg tcctcacagc tttagaaaat
     1381 ggttccatca gcacctggga tgtagagact cgacagctac tcaggcaaat caccacagcc
     1441 cagtctgtca tcctgggcat gaaactcacc agtgatgaaa agtaccttgt ggtggctaca
     1501 acaaataaca ccttgttgat ttatgacaat gtcaattctt gcctcctgtc tgaagtagaa
     1561 atcaagggga ccaagcatgg aagcagcgcc acctacatca atggatttac actgtccgcc
     1621 aaccacgccc ttgcatggct cgaagccagc aaagatgtca ctgtcatcga tctgctgtac
     1681 ggatggccgc tttaccagtt ccactgctgg tatgaagtga cgtgcgtcca gtgctccctg
     1741 gatggtctgt atgctttctg tggccaatac ctgaacacaa ccaccatatt tcatttaggg
     1801 agtggagaaa agttatgtac agtgacatca gaattttcag gtggatttgt gaagtttctt
     1861 cttatcttgg acacagctca ggaaatggtc atggtagaca gtgaaggaag tctttctgtt
     1921 tggaatactg aggacatttc cagcccccag ctgactgatg actttgattg ccgaagagaa
     1981 gacagtgagg tggtcagcat tgagctttca gaagaccaaa gtgcagttct gatctgtaaa
     2041 gccctcagca ttgagctctt agatactggt ctgtggaagg tggctgaaaa gttcagagcc
     2101 aagcacaacg aacgctttat atctgccgtg ctgtctaaaa atggagattg tatcatcgcc
     2161 accatggaaa atacctcagc tgtgtttttt tggaggcggg acacaggaca gtgtatggca
     2221 agcttgcagg aaatctcagg ttccattgtt aagttagtga aatccagtca ccacaatatg
     2281 ctactgtctt tatcaaccag tggtgttctt tccatttggg acatagatat aatcacagct
     2341 atgtccaaca tagataagac tggaaaaccc atccaaagtc tgttgttgcc tgctagaggg
     2401 gaaatcattt actccctgga tggatccgat tgtgttcata agtggaactt cagcagtggc
     2461 ttcatcgaag cagtattcaa gcatgaagga atagttgaac actgtgtgtt aacatccacc
     2521 ggagatataa tggtgacatc agatgacaaa agcagccagt atgtctggca caccagcagt
     2581 ggtgaaaacc tttttcgaat taacgggcag agaatatctc agctgctgat tacacacaat
     2641 gaccagtttg tggtctcgct ctgtgaggaa aatgcctcca gggtttggag gctcgccaca
     2701 ggccacaggg tctgcaacat tctgaccact ttgcagaatg cctttattac ctccgcaaat
     2761 accttcgtgg tgggaatgac taaaagcaaa gtgttggcag tcagtctttg gacaggaagt
     2821 atcaccaaga aattttgctg tgaagatggg accaccatcg tgaattttaa attaatccct
     2881 gactgtcctg atatcatcgt gtttatcaca tcggccgaga ctgtgaacat ctggagtctg
     2941 acagatgaag tgatctgtcg gcgcgtgcaa cttccaaaca acttcttgaa aaatctggag
     3001 gactttgaaa tttctcccaa tggaaagcta ggcattatag ccaggggaga tgaaaacatc
     3061 aatgtgctag atttatacag tggtaaattg cgggtggttc acgcctctgg gatcatctgg
     3121 cggcagaggt tgtctcggga tggtcgctac ctggtataca tttgtttccg aaatggggag
     3181 gaggaggatg aaaatggtgc aatattcagt ttaattgtaa tgagactggc agatggcaaa
     3241 aatatcggtg cttgttccct ttacaaaaca ccaactttcc ttgcactctc ccagaggcac
     3301 ctgaacatca ttgtgggctt tgatgatggg agtataggga tctacacggt agtagaccgt
     3361 gtagatgctg cactgaaaat caaaattgcc acttcaaata gcggctcgag cggtgggagc
     3421 tccggtgtga gcggctggcg gctgttcaag aagattagct aagtttaaac ggccgcgact
     3481 ctagactgca ggcatgcaag ctgatttggc tgctgccacc gctgagcaag gccgcttcga
     3541 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3601 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3661 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     3721 tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatctt aattaagact
     3781 tctaatataa cttcgtatag catacattat acgaagttat ggttccactg ttgacaatta
     3841 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga
     3901 ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3961 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4021 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4081 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4141 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4201 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4261 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4321 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4381 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4441 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4501 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4561 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4621 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4681 gccttcttga cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc
     4741 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4801 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4861 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4921 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4981 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5041 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5101 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5161 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5221 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5281 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5341 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5401 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     5461 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5521 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca
     5581 aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc
     5641 tagagtttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt
     5701 cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac
     5761 ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat
     5821 acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga gttctgcaca
     5881 aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag
     5941 agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc
     6001 tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatggtgg cgctagtgtc
     6061 agaagaatcg agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     6121 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     6181 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     6241 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     6301 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     6361 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     6421 ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt caggccatgg
     6481 tgctgcgc
//
