LOCUS       sequence-423401-        6342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human UTP4. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6342)
  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 6342)
  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_UTP4:1-686
FEATURES             Location/Qualifiers
     source          1..6342
                     /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    3405..3526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3416..3435)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3641..3674)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3684..3731
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3763..4557
                     /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(3817..3836)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4427..4446
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4715..5303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4795..4814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5429..5477
                     /note="synthetic polyadenylation signal"
     CDS             complement(5501..5899)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5896..5905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5930..6287)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5944..6079
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6096..6287
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6244..6264
                     /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 tgggtgaatt
     1261 taaggtccat cgagtacgtt tctttaatta tgttccatca ggaatccgct gtgtggctta
     1321 caataaccag tcaaacagat tggctgtttc acgaacagat ggcactgtgg aaatttataa
     1381 cttgtcagca aactactttc aggagaaatt tttcccaggt catgagtctc gggctacaga
     1441 agctttgtgc tgggcagaag gacagcgact ctttagtgct gggctcaatg gcgagattat
     1501 ggagtatgat ttacaggcgt taaacatcaa gtatgctatg gatgcctttg gaggacctat
     1561 ttggagcatg gctgccagcc ccagtggctc tcaacttttg gttggttgtg aagatggatc
     1621 tgtgaaacta tttcaaatta ccccagacaa aatccagttt gaaagaaatt ttgatcggca
     1681 gaaaagtcgc atcctgagtc tcagctggca tccctctggt acccacattg cagctggttc
     1741 catagactac attagtgtgt ttgatgtcaa atcaggcagc gctgttcata agatgattgt
     1801 ggacaggcag tatatgggcg tgtctaagcg gaagtgcatc gtgtggggtg tcgccttctt
     1861 gtccgatggc actatcataa gtgtggactc tgctgggaag gtgcagttct gggactcagc
     1921 cactgggacg cttgtgaaga gccatctcat cgctaatgct gacgtgcagt ccattgctgt
     1981 agctgaccaa gaagacagtt tcgtggtggg cacagccgag ggaacagtct tccattttca
     2041 gctggtccct gtgacatcta acagcagtga gaagcagtgg gtgcggacaa aaccgttcca
     2101 gcatcacact catgacgtgc gcactgtggc ccacagccca acagcgctga tatctggagg
     2161 cactgacacc cacttagtct ttcgtcctct catggagaag gtggaagtaa agaattacga
     2221 tgccgctctc cgaaaaatca cctttcccca ccgatgtctc atctcctgtt ctaaaaagag
     2281 gcagcttctc ctcttccagt ttgctcatca cttagaactt tggcgactgg gatccacagt
     2341 tgcaacaggc aagaatgggg atactcttcc actctctaaa aatgcagatc atttactgca
     2401 cctaaagaca aagggtcctg agaacattat ctgtagctgt atctccccat gtggaagttg
     2461 gatagcctat tctacagttt ctcggttttt tctctatcgg ctgaattatg aacatgacaa
     2521 cataagcctc aaaagggttt ccaaaatgcc agcattcctt cgctctgccc ttcagatttt
     2581 gttttctgaa gattcaacaa agctctttgt agcatcaaat caaggagctc tgcatattgt
     2641 tcagctgtca ggaggaagct tcaagcacct gcatgctttc cagcctcagt caggaacagt
     2701 ggaggccatg tgtcttttgg cagtcagtcc agatgggaat tggctagctg catcaggtac
     2761 cagtgctgga gtccatgtct acaacgtaaa acagctaaag cttcactgca cggtgcctgc
     2821 ttacaatttc ccagtgactg ctatggctat tgcccccaat accaacaacc ttgtcatcgc
     2881 tcattcggac cagcaggtat ttgagtacag catcccagac aaacagtata cagattggag
     2941 ccggactgtc cagaagcagg gctttcacca cctttggctc caaagggata ctcctatcac
     3001 acacatcagt tttcatccca agagaccgat gcacatcctt ctccatgatg cctacatgtt
     3061 ctgcatcatt gacaagtcat tgccccttcc aaatgacaaa accttactct acaatccatt
     3121 tcctcccacg aatgaatcag atgtcatccg gaggcgcaca gctcatgctt ttaaaatttc
     3181 taagatatat aagcctctac tcttcatgga tcttttggat gaaagaacac tcgtggcagt
     3241 agaacggcct ctggatgaca tcattgctca gctcccacca cccattaaaa agaagaaatt
     3301 tggaacctaa tctagaggtc aagcttcggg ggtttaaacc tgcaggcatg caagctgatt
     3361 tggctgctgc caccgctgag caaggccgct tcgagcagac atgataagat acattgatga
     3421 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3481 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3541 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     3601 cctctacaaa tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca
     3661 ttatacgaag ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag
     3721 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     3781 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3841 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3901 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3961 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4021 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4081 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4141 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4201 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4261 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4321 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4381 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4441 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4501 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc
     4561 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     4621 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4681 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4741 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4801 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4861 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4921 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4981 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5041 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5101 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5161 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5221 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5281 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5341 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5401 cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa
     5461 aataaagata ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac
     5521 cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta
     5581 tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca
     5641 agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag
     5701 cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat
     5761 tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt
     5821 cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt
     5881 gagacaaagg cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc
     5941 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     6001 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     6061 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     6121 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     6181 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     6241 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc
     6301 aagttcctct ttcagaggtt atttcaggcc atggtgctgc gc
//
