LOCUS       sequence-423453-        5972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WRAP53. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5972)
  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 5972)
  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_WRAP53:1-548
FEATURES             Location/Qualifiers
     source          1..5972
                     /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    3035..3156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3046..3065)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3271..3304)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3314..3361
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3393..4187
                     /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(3447..3466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4057..4076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4345..4933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4425..4444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5059..5107
                     /note="synthetic polyadenylation signal"
     CDS             complement(5131..5529)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5526..5535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5560..5917)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5574..5709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5726..5917
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5874..5894
                     /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 ccatgaagac tttggagact
     1261 caaccgttag ctccggactg ctgtccttca gaccaggacc cagctccagc ccatccttct
     1321 ccccacgctt ccccgatgaa taaaaatgcg gactctgaac tgatgccacc gcctcccgaa
     1381 aggggggatc cgccccggtt gtccccagat cctgtggctg gctcagctgt gtcccaggag
     1441 ctacgggagg gggacccagt ttctctctcc actcccctgg aaacagagtt tggttcccct
     1501 agtgagttga gtcctcgaat cgaggagcaa gaactttctg aaaatacaag ccttcctgca
     1561 gaagaagcaa acgggagcct ttctgaagaa gaagcgaacg ggccagagtt ggggtctgga
     1621 aaagccatgg aagatacctc tggggaaccc gctgcagagg acgagggaga caccgcttgg
     1681 aactacagct tctcccagct gcctcgattt ctcagtggtt cctggtcaga gttcagcacc
     1741 caacctgaga acttcttgaa aggctgtaag tgggctcctg acggttcctg catcttgacc
     1801 aatagtgctg ataacatctt gcgaatttat aacctgcccc cagagctgta ccatgagggg
     1861 gagcaggtgg aatatgcaga aatggtccct gtccttcgaa tggtggaagg tgataccatc
     1921 tatgattact gctggtattc tctgatgtcc tcagcccagc cagacacctc ctacgtggcc
     1981 agcagcagcc gggagaaccc gattcatatc tgggacgcat tcactggaga gctccgggct
     2041 tcctttcgcg cctacaacca cctggatgag ctgacggcag cccattcgct ctgcttctcc
     2101 ccggatggct cccagctctt ctgtggcttc aaccggactg tgcgtgtttt ttccacggcc
     2161 cggcctggcc gagactgcga ggtccgagcc acatttgcaa aaaagcaggg ccagagcggc
     2221 atcatctcct gcatagcctt cagcccagcc cagcccctct atgcctgtgg ctcctacggc
     2281 cgctccctgg gtctgtatgc ctgggatgat ggctcccctc tcgccttgct gggagggcac
     2341 caagggggca tcacccacct ctgctttcat cccgatggca accgcttctt ctcaggagcc
     2401 cgcaaggatg ctgagctcct gtgctgggat ctccggcagt ctggttaccc actgtggtcc
     2461 ctgggtcgag aggtgaccac caatcagcgc atctacttcg atctggaccc gaccgggcag
     2521 ttcctagtga gtggcagcac gagcggggct gtctctgtgt gggacacgga cgggcctggc
     2581 aatgatggga agccggagcc cgtgttgagt tttctgcccc agaaggactg caccaatggc
     2641 gtgagcctgc accctagcct gcctctcctg gccactgcct ccggtcagcg tgtgtttcct
     2701 gagcccacag agagtgggga cgaaggagag gagctgggcc ttcccttgct ctccacgcgc
     2761 cacgtccacc ttgaatgtcg gcttcagctc tggtggtgtg ggggggcgcc agactccagc
     2821 atccctgatg atcaccaggg cgagaaaggg cagggaggaa cggagggagg tgtgggtgag
     2881 ctgataggct cgagcggtgg gagctccggt gtgagcggct ggcggctgtt caagaagatt
     2941 agctaagttt aaacggccgc gactctagac tgcaggcatg caagctgatt tggctgctgc
     3001 caccgctgag caaggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     3061 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     3121 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3181 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     3241 tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca ttatacgaag
     3301 ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3361 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     3421 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3481 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3541 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3601 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3661 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3721 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3781 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3841 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3901 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3961 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4021 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4081 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4141 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaatgaccg
     4201 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4261 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4321 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4381 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4441 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4501 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4561 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4621 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4681 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4741 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     4801 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4861 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4921 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4981 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5041 tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa aataaagata
     5101 ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac cagagggcag
     5161 caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta tggctttgat
     5221 cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca agatgcccct
     5281 gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag cagcagcagt
     5341 gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat tgacaccagt
     5401 gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt cagagatggg
     5461 gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt gagacaaagg
     5521 cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     5581 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     5641 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     5701 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     5761 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     5821 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     5881 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct
     5941 ttcagaggtt atttcaggcc atggtgctgc gc
//
