LOCUS       sequence-425798-        5772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FZR1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5772)
  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 5772)
  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_FZR1:1-496
FEATURES             Location/Qualifiers
     source          1..5772
                     /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(1835..1846)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2835..2956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2846..2865)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3071..3104)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3114..3161
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3193..3987
                     /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(3247..3266)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3857..3876
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4145..4733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4225..4244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4859..4907
                     /note="synthetic polyadenylation signal"
     CDS             complement(4931..5329)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5326..5335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5360..5717)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5374..5509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5526..5717
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5674..5694
                     /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 tggaccagga
     1261 ctatgagcgg cgcctgcttc gccagatcgt catccagaat gagaacacga tgccacgcgt
     1321 cacagagatg cggcggaccc tgacgcctgc cagctcccca gtgtcctcgc ccagcaagca
     1381 cggagaccgc ttcatcccct ccagagccgg agccaactgg agcgtgaact tccacaggat
     1441 taacgagaat gagaagtctc ccagtcagaa ccggaaagcc aaggacgcta cctcagacaa
     1501 cggcaaagac ggcctggcct actctgccct gctcaagaat gagctgctgg gtgccggcat
     1561 cgagaaggtg caggacccgc agactgagga ccgcaggctg cagccctcca cgcctgagaa
     1621 gaagggtctg ttcacgtatt cccttagcac caagcgctcc agccccgatg acggcaacga
     1681 tgtgtctccc tactccctgt ctcccgtcag caacaagagc cagaagctgc tccggtcccc
     1741 ccggaaaccc acccgcaaga tctccaagat ccccttcaag gtgctggacg cgcccgagct
     1801 gcaggacgac ttctacctca atctggtgga ctggtcgtcc ctcaatgtgc tcagcgtggg
     1861 gctaggcacc tgcgtgtacc tgtggagtgc ctgtaccagc caggtgacgc ggctctgtga
     1921 cctctcagtg gaaggggact cagtgacctc cgtgggctgg tctgagcggg ggaacctggt
     1981 ggcggtgggc acacacaagg gcttcgtgca gatctgggac gcagccgcag ggaagaagct
     2041 gtccatgttg gagggccaca cggcacgcgt cggggcgctg gcctggaatg ctgagcagct
     2101 gtcgtccggg agccgcgacc gcatgatcct gcagagggac atccgcaccc cgccactgca
     2161 gtcggagcgg cggctgcagg gccaccggca ggaggtgtgc gggctcaagt ggtccacaga
     2221 ccaccagctc ctcgcctcgg ggggcaacga caacaagctg ctggtctgga atcactcgag
     2281 cctgagcccc gtgcagcagt acacggagca cctggcggcc gtgaaggcca tcgcctggtc
     2341 cccacatcag cacgggctgc tggcctcggg gggcggcaca gctgaccgct gtatccgctt
     2401 ctggaacacg ctgacaggac aaccactgca gtgtatcgac acgggctccc aagtgtgcaa
     2461 tctggcctgg tccaagcacg ccaacgagct ggtgagcacg cacggctact cacagaacca
     2521 gatccttgtc tggaagtacc cctccctgac ccaggtggcc aagctgaccg ggcactccta
     2581 ccgcgtgctg tacctggcaa tgtcccctga tggggaggcc atcgtcactg gtgctggaga
     2641 cgagaccctg aggttctgga acgtctttag caaaacccgt tcgacaaagg taaagtggga
     2701 gtctgtgtct gtgctcaacc tcttcaccag gatccggtaa tctagaggtc aagcttcggg
     2761 ggtttaaacc tgcaggcatg caagctgatt tggctgctgc caccgctgag caaggccgct
     2821 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2881 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2941 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3001 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa
     3061 gacttctaat ataacttcgt atagcataca ttatacgaag ttatggttcc actgttgaca
     3121 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     3181 aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3241 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3301 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3361 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3421 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3481 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3541 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3601 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3661 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3721 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3781 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3841 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3901 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3961 tatcgccttc ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg
     4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4321 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4561 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4801 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca
     4861 cacaaaaaac caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga
     4921 tggctagagt ttagccctcc cacacataac cagagggcag caattcacga atcccaactg
     4981 ccgtcggctg tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa
     5041 gcacctgtcg gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga
     5101 cgatacaagt caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg
     5161 cacaaggtcc cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag
     5221 agagagctgc gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg
     5281 ttgctctttc aatgagggtg gattcttctt gagacaaagg cttggccatg gtggcgctag
     5341 tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     5401 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     5461 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     5521 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5581 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     5641 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
     5701 ggttctttcc gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc
     5761 atggtgctgc gc
//
