LOCUS       sequence-423355-        6023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human USP21. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6023)
  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 6023)
  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_USP21:1-565
FEATURES             Location/Qualifiers
     source          1..6023
                     /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    3086..3207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3097..3116)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3322..3355)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3365..3412
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3444..4238
                     /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(3498..3517)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4108..4127
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4396..4984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4476..4495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5110..5158
                     /note="synthetic polyadenylation signal"
     CDS             complement(5182..5580)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5577..5586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5611..5968)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5625..5760
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5777..5968
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5925..5945
                     /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 ccatgcccca ggcctctgag
     1261 caccgcctgg gccgtacccg agagccacct gttaatatcc agccccgagt gggatccaag
     1321 ctaccatttg cccccagggc ccgcagcaag gagcgcagaa acccagcctc tgggccaaac
     1381 cccatgttac gacctctgcc tccccggcca ggtctgcctg atgaacggct caagaaactg
     1441 gagctgggac ggggacggac ctcaggccct cgtcccagag gcccccttcg agcagatcat
     1501 ggggttcccc tgcctggctc accaccccca acagtggctt tgcctctccc atctcggacc
     1561 aacttagccc gttccaagtc tgtgagcagt ggggacttgc gtccaatggg gattgccttg
     1621 ggagggcacc gtggcaccgg agagcttggg gctgcactga gccgcttggc cctccggcct
     1681 gagccaccca ctttgagacg tagcacttct ctccgccgcc tagggggctt tcctggaccc
     1741 cctaccctgt tcagcatacg gacagagccc cctgcttccc atggctcctt ccacatgata
     1801 tccgcccggt cctctgagcc tttctactct gatgacaaga tggctcatca cacactcctt
     1861 ctgggctctg gtcatgttgg ccttcgaaac ctgggaaaca cgtgcttcct gaatgctgtg
     1921 ctgcagtgtc tgagcagcac tcgacctctt cgggacttct gtctgagaag ggacttccgg
     1981 caagaggtgc ctggaggagg ccgagcccaa gagctcactg aagcctttgc agatgtgatt
     2041 ggtgccctct ggcaccctga ctcctgcgaa gctgtgaatc ctactcgatt ccgagctgtc
     2101 ttccagaaat atgttccctc cttctctgga tacagccagc aggatgccca agagttcctg
     2161 aagctcctca tggagcggct acaccttgaa atcaaccgcc gaggccgccg ggctccaccg
     2221 atacttgcca atggtccagt tccctctcca ccccgccgag gaggggctct gctagaagaa
     2281 cctgagttaa gtgatgatga ccgagccaac ctaatgtgga aacgttacct ggagcgagag
     2341 gacagcaaga ttgtggacct gtttgtgggc cagttgaaaa gttgtctcaa gtgccaggcc
     2401 tgtgggtatc gctccacgac cttcgaggtt ttttgtgacc tgtccctgcc catccccaag
     2461 aaaggatttg ctgggggcaa ggtgtctctg cgggattgtt tcaacctttt cactaaggaa
     2521 gaagagctag agtcggagaa tgccccagtg tgtgaccgat gtcggcagaa aactcgaagt
     2581 accaaaaagt tgacagtaca aagattccct cgaatcctcg tgctccatct gaatcgattt
     2641 tctgcctccc gaggctccat caaaaaaagt tcagtaggtg tagactttcc actgcagcga
     2701 ctgagcctag gggactttgc cagtgacaaa gccggaagtc ctgtatacca gctgtatgcc
     2761 ctttgcaacc actcaggcag cgtccactat ggccactaca cagccctgtg ccggtgccag
     2821 actggttggc atgtctacaa tgactctcgt gtctcccctg tcagtgaaaa ccaggtggca
     2881 tccagcgagg gctacgtgct gttctaccaa ctgatgcagg agccaccccg gtgcctgggc
     2941 tcgagcggtg ggagctccgg tgtgagcggc tggcggctgt tcaagaagat tagctaagtt
     3001 taaacggccg cgactctaga ctgcaggcat gcaagctgat ttggctgctg ccaccgctga
     3061 gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3121 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3181 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3241 gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa
     3301 atcttaatta agacttctaa tataacttcg tatagcatac attatacgaa gttatggttc
     3361 cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     3421 gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg
     3481 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3541 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3601 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3661 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3721 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3781 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3841 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3901 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3961 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4021 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4081 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4141 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4201 gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc gaccaagcga
     4261 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4321 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4381 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4441 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4501 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4561 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4621 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4681 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4741 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4801 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4861 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4921 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     4981 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5041 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     5101 cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     5161 ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca gcaattcacg
     5221 aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga tcccaggatg
     5281 cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt
     5341 tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag tgcccagcac
     5401 cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag tgaagatgcg
     5461 gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg ggatgctgtt
     5521 gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag gcttggccat
     5581 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     5641 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     5701 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     5761 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     5821 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     5881 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     5941 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     6001 tatttcaggc catggtgctg cgc
//
