LOCUS       sequence-425869-        5816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FZR1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5816)
  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 5816)
  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_FZR1:1-496
FEATURES             Location/Qualifiers
     source          1..5816
                     /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"
     CDS             complement(1828..1839)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2879..3000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2890..2909)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3115..3148)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3158..3205
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3237..4031
                     /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(3291..3310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3901..3920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4189..4777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4269..4288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4903..4951
                     /note="synthetic polyadenylation signal"
     CDS             complement(4975..5373)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5370..5379
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5404..5761)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5418..5553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5570..5761
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5718..5738
                     /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 ccatggacca ggactatgag
     1261 cggcgcctgc ttcgccagat cgtcatccag aatgagaaca cgatgccacg cgtcacagag
     1321 atgcggcgga ccctgacgcc tgccagctcc ccagtgtcct cgcccagcaa gcacggagac
     1381 cgcttcatcc cctccagagc cggagccaac tggagcgtga acttccacag gattaacgag
     1441 aatgagaagt ctcccagtca gaaccggaaa gccaaggacg ccacctcaga caacggcaaa
     1501 gacggcctgg cctactctgc cctgctcaag aatgagctgc tgggtgccgg catcgagaag
     1561 gtgcaggacc cgcagactga ggaccgcagg ctgcagccct ccacgcctga gaagaagggt
     1621 ctgttcacgt attcccttag caccaagcgc tccagccccg atgacggcaa cgatgtgtct
     1681 ccctactccc tgtctcccgt cagcaacaag agccagaagc tgctccggtc cccccggaaa
     1741 cccacccgca agatctccaa gatccccttc aaggtgctgg acgcgcccga gctgcaggac
     1801 gacttctacc tcaatctggt ggactggtcg tccctcaatg tgctcagcgt ggggctaggc
     1861 acctgcgtgt acctgtggag tgcctgtacc agccaggtga cgcggctctg tgacctctca
     1921 gtggaagggg actcagtgac ctccgtgggc tggtctgagc gggggaacct ggtggcggtg
     1981 ggcacacaca agggcttcgt gcagatctgg gacgcagccg cagggaagaa gctgtccatg
     2041 ttggagggcc acacggcacg cgtcggggcg ctggcctgga atgctgagca gctgtcgtcc
     2101 gggagccgcg accgcatgat cctgcagagg gacatccgca ccccgccact gcagtcggag
     2161 cggcggctgc agggccaccg gcaggaggtg tgcgggctca agtggtccac agaccaccag
     2221 ctcctcgcct cggggggcaa cgacaacaag ctgctggtct ggaatcactc gagcctgagc
     2281 cccgtgcagc agtacacgga gcacctggcg gccgtgaagg ccatcgcctg gtccccacat
     2341 cagcacgggc tgctggcctc ggggggcggc acagctgacc gctgtatccg cttctggaac
     2401 acgctgacag gacaaccact gcagtgtatc gacacgggct cccaagtgtg caatctggcc
     2461 tggtccaagc acgccaacga gctggtgagc acgcacggct actcacagaa ccagatcctt
     2521 gtctggaagt acccctccct gacccaggtg gccaagctga ccgggcactc ctaccgcgtg
     2581 ctgtacctgg caatgtcccc tgatggggag gccatcgtca ctggtgctgg agacgagacc
     2641 ctgaggttct ggaacgtctt tagcaaaacc cgttcgacaa aggtaaagtg ggagtctgtg
     2701 tctgtgctca acctcttcac caggatccgg ggctcgagcg gtgggagctc cggtgtgagc
     2761 ggctggcggc tgttcaagaa gattagctaa gtttaaacgg ccgcgactct agactgcagg
     2821 catgcaagct gatttggctg ctgccaccgc tgagcaaggc cgcttcgagc agacatgata
     2881 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2941 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3001 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     3061 ttaagcaagt aaaacctcta caaatgtggt aaaatcttaa ttaagacttc taatataact
     3121 tcgtatagca tacattatac gaagttatgg ttccactgtt gacaattaat catcggcata
     3181 gtatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatga
     3241 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3301 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3361 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3421 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3481 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3541 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3601 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3661 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3721 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3781 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3841 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3901 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3961 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4021 agttcttctg attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa
     4081 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4141 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4201 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4261 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4321 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4381 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4441 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4501 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4561 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4621 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4681 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4741 caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac
     4801 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4861 aaaaaggatc ttcacctaga tccttttata gtccggaatc gtcacacaaa aaaccaacac
     4921 acagatgtaa tgaaaataaa gatattttat tgcggccatc gtgatggcta gagtttagcc
     4981 ctcccacaca taaccagagg gcagcaattc acgaatccca actgccgtcg gctgtccatc
     5041 actgtccttc actatggctt tgatcccagg atgcagatcg agaagcacct gtcggcaccg
     5101 tccgcagggg ctcaagatgc ccctgttctc atttccgatc gcgacgatac aagtcaggtt
     5161 gccagctgcc gcagcagcag cagtgcccag caccacgagt tctgcacaag gtcccccagt
     5221 aaaatgatat acattgacac cagtgaagat gcggccgtcg ctagagagag ctgcgctggc
     5281 gacgctgtag tcttcagaga tggggatgct gttgattgta gccgttgctc tttcaatgag
     5341 ggtggattct tcttgagaca aaggcttggc catggtggcg ctagtgtcag aagaatcgag
     5401 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     5461 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     5521 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5581 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5641 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     5701 gcctggggac tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca
     5761 gaaggtacct aaccaagttc ctctttcaga ggttatttca ggccatggtg ctgcgc
//
