LOCUS       sequence-425926-        5928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WRAP53. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5928)
  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 5928)
  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_WRAP53:1-548
FEATURES             Location/Qualifiers
     source          1..5928
                     /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    2991..3112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3002..3021)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3227..3260)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3270..3317
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3349..4143
                     /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(3403..3422)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4013..4032
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4301..4889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4381..4400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5015..5063
                     /note="synthetic polyadenylation signal"
     CDS             complement(5087..5485)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5482..5491
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5516..5873)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5530..5665
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5682..5873
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5830..5850
                     /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 tgaagacttt
     1261 ggagactcaa ccgttagctc cggactgctg tccttcagac caggacccag ctccagccca
     1321 tccttctccc cacgcttccc cgatgaataa aaatgcggac tctgaactga tgccaccgcc
     1381 tcccgaaagg ggggatccgc cccggttgtc cccagatcct gtggctggct cagctgtgtc
     1441 ccaggagcta cgggaggggg acccagtttc tctctccact cccctggaaa cagagtttgg
     1501 ttcccctagt gagttgagtc ctcgaatcga ggagcaagaa ctttctgaaa atacaagcct
     1561 tcctgcagaa gaagcaaacg ggagcctttc tgaagaagaa gcgaacgggc cagagttggg
     1621 gtctggaaaa gccatggaag atacctctgg ggaacccgct gcagaggacg agggagacac
     1681 cgcttggaac tacagcttct cccagctgcc tcgatttctc agtggttcct ggtcagagtt
     1741 cagcacccaa cctgagaact tcttgaaagg ctgtaagtgg gctcctgacg gttcctgcat
     1801 cttgaccaat agtgctgata acatcttgcg aatttataac ctgcccccag agctgtacca
     1861 tgagggggag caggtggaat atgcagaaat ggtccctgtc cttcgaatgg tggaaggtga
     1921 taccatctat gattactgct ggtattctct gatgtcctca gcccagccag acacctccta
     1981 cgtggccagc agcagccggg agaacccgat tcatatctgg gacgcattca ctggagagct
     2041 ccgggcttcc tttcgcgcct acaaccacct ggatgagctg acggcagccc attcgctctg
     2101 cttctccccg gatggctccc agctcttctg tggcttcaac cggactgtgc gtgttttttc
     2161 cacggcccgg cctggccgag actgcgaggt ccgagccaca tttgcaaaaa agcagggcca
     2221 gagcggcatc atctcctgca tagccttcag cccagcccag cccctctatg cctgtggctc
     2281 ctacggccgc tccctgggtc tgtatgcctg ggatgatggc tcccctctcg ccttgctggg
     2341 agggcaccaa gggggcatca cccacctctg ctttcatccc gatggcaacc gcttcttctc
     2401 aggagcccgc aaggatgctg agctcctgtg ctgggatctc cggcagtctg gttacccact
     2461 gtggtccctg ggtcgagagg tgaccaccaa tcagcgcatc tacttcgatc tggacccgac
     2521 cgggcagttc ctagtgagtg gcagcacgag cggggctgtc tctgtgtggg acacggacgg
     2581 gcctggcaat gatgggaagc cggagcccgt gttgagtttt ctgccccaga aggactgcac
     2641 caatggcgtg agcctgcacc ctagcctgcc tctcctggcc actgcctccg gtcagcgtgt
     2701 gtttcctgag cccacagaga gtggggacga aggagaggag ctgggccttc ccttgctctc
     2761 cacgcgccac gtccaccttg aatgtcggct tcagctctgg tggtgtgggg gggcgccaga
     2821 ctccagcatc cctgatgatc accagggcga gaaagggcag ggaggaacgg agggaggtgt
     2881 gggtgagctg atataatcta gaggtcaagc ttcgggggtt taaacctgca ggcatgcaag
     2941 ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat
     3001 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3061 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3121 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     3181 gtaaaacctc tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag
     3241 catacattat acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg
     3301 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     3361 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3421 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3481 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3541 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3601 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3661 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3721 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3781 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3841 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3901 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3961 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4021 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4081 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4141 tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     4201 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4321 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4741 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4861 agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg
     4921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4981 tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt
     5041 aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca
     5101 cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct
     5161 tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg
     5221 ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg
     5281 ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat
     5341 atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt
     5401 agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt
     5461 cttcttgaga caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc
     5521 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     5581 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     5641 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     5701 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     5761 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5821 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac
     5881 ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgc
//
