LOCUS       sequence-423382-        5979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human USP21. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5979)
  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 5979)
  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_USP21:1-565
FEATURES             Location/Qualifiers
     source          1..5979
                     /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    3042..3163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3053..3072)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3278..3311)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3321..3368
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3400..4194
                     /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(3454..3473)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4064..4083
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4352..4940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4432..4451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5066..5114
                     /note="synthetic polyadenylation signal"
     CDS             complement(5138..5536)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5533..5542
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5567..5924)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5581..5716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5733..5924
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5881..5901
                     /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 tgccccaggc
     1261 ctctgagcac cgcctgggcc gtacccgaga gccacctgtt aatatccagc cccgagtggg
     1321 atccaagcta ccatttgccc ccagggcccg cagcaaggag cgcagaaacc cagcctctgg
     1381 gccaaacccc atgttacgac ctctgcctcc ccggccaggt ctgcctgatg aacggctcaa
     1441 gaaactggag ctgggacggg gacggacctc aggccctcgt cccagaggcc cccttcgagc
     1501 agatcatggg gttcccctgc ctggctcacc acccccaaca gtggctttgc ctctcccatc
     1561 tcggaccaac ttagcccgtt ccaagtctgt gagcagtggg gacttgcgtc caatggggat
     1621 tgccttggga gggcaccgtg gcaccggaga gcttggggct gcactgagcc gcttggccct
     1681 ccggcctgag ccacccactt tgagacgtag cacttctctc cgccgcctag ggggctttcc
     1741 tggaccccct accctgttca gcatacggac agagccccct gcttcccatg gctccttcca
     1801 catgatatcc gcccggtcct ctgagccttt ctactctgat gacaagatgg ctcatcacac
     1861 actccttctg ggctctggtc atgttggcct tcgaaacctg ggaaacacgt gcttcctgaa
     1921 tgctgtgctg cagtgtctga gcagcactcg acctcttcgg gacttctgtc tgagaaggga
     1981 cttccggcaa gaggtgcctg gaggaggccg agcccaagag ctcactgaag cctttgcaga
     2041 tgtgattggt gccctctggc accctgactc ctgcgaagct gtgaatccta ctcgattccg
     2101 agctgtcttc cagaaatatg ttccctcctt ctctggatac agccagcagg atgcccaaga
     2161 gttcctgaag ctcctcatgg agcggctaca ccttgaaatc aaccgccgag gccgccgggc
     2221 tccaccgata cttgccaatg gtccagttcc ctctccaccc cgccgaggag gggctctgct
     2281 agaagaacct gagttaagtg atgatgaccg agccaaccta atgtggaaac gttacctgga
     2341 gcgagaggac agcaagattg tggacctgtt tgtgggccag ttgaaaagtt gtctcaagtg
     2401 ccaggcctgt gggtatcgct ccacgacctt cgaggttttt tgtgacctgt ccctgcccat
     2461 ccccaagaaa ggatttgctg ggggcaaggt gtctctgcgg gattgtttca accttttcac
     2521 taaggaagaa gagctagagt cggagaatgc cccagtgtgt gaccgatgtc ggcagaaaac
     2581 tcgaagtacc aaaaagttga cagtacaaag attccctcga atcctcgtgc tccatctgaa
     2641 tcgattttct gcctcccgag gctccatcaa aaaaagttca gtaggtgtag actttccact
     2701 gcagcgactg agcctagggg actttgccag tgacaaagcc ggaagtcctg tataccagct
     2761 gtatgccctt tgcaaccact caggcagcgt ccactatggc cactacacag ccctgtgccg
     2821 gtgccagact ggttggcatg tctacaatga ctctcgtgtc tcccctgtca gtgaaaacca
     2881 ggtggcatcc agcgagggct acgtgctgtt ctaccaactg atgcaggagc caccccggtg
     2941 cctgtaatct agaggtcaag cttcgggggt ttaaacctgc aggcatgcaa gctgatttgg
     3001 ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3061 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3121 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3181 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3241 ctacaaatgt ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta
     3301 tacgaagtta tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     3361 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     3421 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3481 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3541 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3601 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3661 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3721 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3781 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3841 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3901 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3961 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4021 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4081 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4141 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa
     4201 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     4261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4921 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5041 agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     5101 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     5161 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     5221 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     5281 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     5341 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     5401 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     5461 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     5521 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     5581 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     5641 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     5701 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     5761 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     5821 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5881 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     5941 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
