LOCUS       sequence-423468-        6386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human UTP4. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  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 6386)
  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_UTP4:1-686
FEATURES             Location/Qualifiers
     source          1..6386
                     /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    3449..3570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3460..3479)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3685..3718)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3728..3775
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3807..4601
                     /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(3861..3880)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4471..4490
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4759..5347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4839..4858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5473..5521
                     /note="synthetic polyadenylation signal"
     CDS             complement(5545..5943)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5940..5949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5974..6331)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5988..6123
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6140..6331
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6288..6308
                     /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 ccatgggtga atttaaggtc
     1261 catcgagtac gtttctttaa ttatgttcca tcaggaatcc gctgtgtggc ttacaataac
     1321 cagtcaaaca gattggctgt ttcacgaaca gatggcactg tggaaattta taacttgtca
     1381 gcaaactact ttcaggagaa atttttccca ggtcatgagt ctcgggctac agaagctttg
     1441 tgctgggcag aaggacagcg actctttagt gctgggctca atggcgagat tatggagtat
     1501 gatttacagg cgttaaacat caagtatgct atggatgcct ttggaggacc tatttggagc
     1561 atggctgcca gccccagtgg ctctcaactt ttggttggtt gtgaagatgg atctgtgaaa
     1621 ctatttcaaa ttaccccaga caaaatccag tttgaaagaa attttgatcg gcagaaaagt
     1681 cgcatcctga gtctcagctg gcatccctct ggtacccaca ttgcagctgg ttccatagac
     1741 tacattagtg tgtttgatgt caaatcaggc agcgctgttc ataagatgat tgtggacagg
     1801 cagtatatgg gcgtgtctaa gcggaagtgc atcgtgtggg gtgtcgcctt cttgtccgat
     1861 ggcactatca taagtgtgga ctctgctggg aaggtgcagt tctgggactc agccactggg
     1921 acgcttgtga agagccatct catcgctaat gctgacgtgc agtccattgc tgtagctgac
     1981 caagaagaca gtttcgtggt gggcacagcc gagggaacag tcttccattt tcagctggtc
     2041 cctgtgacat ctaacagcag tgagaagcag tgggtgcgga caaaaccgtt ccagcatcac
     2101 actcatgacg tgcgcactgt ggcccacagc ccaacagcgc tgatatctgg aggcactgac
     2161 acccacttag tctttcgtcc tctcatggag aaggtggaag taaagaatta cgatgccgct
     2221 ctccgaaaaa tcacctttcc ccaccgatgt ctcatctcct gttctaaaaa gaggcagctt
     2281 ctcctcttcc agtttgctca tcacttagaa ctttggcgac tgggatccac agttgcaaca
     2341 ggcaagaatg gggatactct tccactctct aaaaatgcag atcatttact gcacctaaag
     2401 acaaagggtc ctgagaacat tatctgtagc tgtatctccc catgtggaag ttggatagcc
     2461 tattctacag tttctcggtt ttttctctat cggctgaatt atgaacatga caacataagc
     2521 ctcaaaaggg tttccaaaat gccagcattc cttcgctctg cccttcagat tttgttttct
     2581 gaagattcaa caaagctctt tgtagcatca aatcaaggag ctctgcatat tgttcagctg
     2641 tcaggaggaa gcttcaagca cctgcatgct ttccagcctc agtcaggaac agtggaggcc
     2701 atgtgtcttt tggcagtcag tccagatggg aattggctag ctgcatcagg taccagtgct
     2761 ggagtccatg tctacaacgt aaaacagcta aagcttcact gcacggtgcc tgcttacaat
     2821 ttcccagtga ctgctatggc tattgccccc aataccaaca accttgtcat cgctcattcg
     2881 gaccagcagg tatttgagta cagcatccca gacaaacagt atacagattg gagccggact
     2941 gtccagaagc agggctttca ccacctttgg ctccaaaggg atactcctat cacacacatc
     3001 agttttcatc ccaagagacc gatgcacatc cttctccatg atgcctacat gttctgcatc
     3061 attgacaagt cattgcccct tccaaatgac aaaaccttac tctacaatcc atttcctccc
     3121 acgaatgaat cagatgtcat ccggaggcgc acagctcatg cttttaaaat ttctaagata
     3181 tataagcctc tactcttcat ggatcttttg gatgaaagaa cactcgtggc agtagaacgg
     3241 cctctggatg acatcattgc tcagctccca ccacccatta aaaagaagaa atttggaacc
     3301 ggctcgagcg gtgggagctc cggtgtgagc ggctggcggc tgttcaagaa gattagctaa
     3361 gtttaaacgg ccgcgactct agactgcagg catgcaagct gatttggctg ctgccaccgc
     3421 tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3481 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3541 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3601 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     3661 aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac gaagttatgg
     3721 ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3781 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     3841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3961 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4321 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     4621 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4681 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4741 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4801 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4861 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4921 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4981 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5101 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5161 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5221 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5281 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5341 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5401 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     5461 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     5521 tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg gcagcaattc
     5581 acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg
     5641 atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc
     5701 atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag
     5761 caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat
     5821 gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct
     5881 gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc
     5941 catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa
     6001 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     6061 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     6121 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     6181 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     6241 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     6301 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
     6361 ggttatttca ggccatggtg ctgcgc
//
