LOCUS       sequence-425871-        5579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PREB. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5579)
  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 5579)
  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_PREB:1-417
FEATURES             Location/Qualifiers
     source          1..5579
                     /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    2642..2763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2653..2672)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2878..2911)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2921..2968
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3000..3794
                     /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(3054..3073)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3664..3683
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3952..4540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4032..4051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4666..4714
                     /note="synthetic polyadenylation signal"
     CDS             complement(4738..5136)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5133..5142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5167..5524)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5181..5316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5333..5524
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5481..5501
                     /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 ccatgggccg gcgccgggcg
     1261 ccagagctgt accgggctcc gttcccgttg tacgcgcttc aggtcgaccc cagcactggg
     1321 ctgctcatcg ctgcgggcgg aggaggcgcc gccaagacag gcataaagaa tggcgtgcac
     1381 tttctgcagc tagagctgat taatgggcgc ttgagtgcct ccttgctgca ctcccatgac
     1441 acagagacac gggccaccat gaacttggca ctggctggtg acatccttgc tgcagggcag
     1501 gatgcccact gtcagctcct gcgcttccag gcacatcaac agcagggcaa caaggcagag
     1561 aaggccggtt ccaaggagca ggggcctcga caaaggaagg gagcagcccc agcagagaag
     1621 aaatgtggag cggaaaccca gcacgagggg ctagaactca gggtagagaa tttgcaggcg
     1681 gtgcagacag actttagctc cgatccactg cagaaagttg tgtgcttcaa ccacgataat
     1741 accctgcttg ccactggagg aacagatggc tacgtccgtg tctggaaggt gcccagcctg
     1801 gagaaggttc tggagttcaa agcccacgaa ggggagattg aagacctggc tttagggcct
     1861 gatggcaagt tggtaaccgt gggccgggac cttaaggcct ctgtgtggca gaaggatcag
     1921 ctggtgacac agctgcactg gcaagaaaat ggacccacct tttccagcac accttaccgc
     1981 taccaggcct gcaggtttgg gcaggttcca gaccagcctg ctggcctgcg actcttcaca
     2041 gtgcaaattc cccacaagcg cctgcgccag ccccctccct gctacctcac agcctgggat
     2101 ggctccaact tcttgcccct tcggaccaag tcctgtggcc atgaagtcgt ctcctgcctc
     2161 gatgtcagtg aatccggcac cttcctaggc ctgggcacag tcactggctc tgttgccatc
     2221 tacatagctt tctctctcca gtgcctctac tacgtgaggg aggcccatgg cattgtggtg
     2281 acggatgtgg cctttctacc tgagaagggt cgtggtccag agctccttgg gtcccatgaa
     2341 actgccctgt tctctgtggc tgtggacagt cgttgccagc tgcatctgtt gccctcacgg
     2401 cggagtgttc ctgtgtggct cctgctcctg ctgtgtgtcg ggcttattat tgtgaccatc
     2461 ctgctgctcc agagtgcctt tccaggtttc cttggctcga gcggtgggag ctccggtgtg
     2521 agcggctggc ggctgttcaa gaagattagc taagtttaaa cggccgcgac tctagactgc
     2581 aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg agcagacatg
     2641 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2701 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2761 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2821 tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac ttctaatata
     2881 acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt aatcatcggc
     2941 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     3001 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3061 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3121 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3181 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3241 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3301 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3361 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3421 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3481 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3541 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3601 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3661 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3721 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3781 acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     3841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4381 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4501 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4621 atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac aaaaaaccaa
     4681 cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagttta
     4741 gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc
     4801 atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca
     4861 ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag
     4921 gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc
     4981 agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct
     5041 ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat
     5101 gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt cagaagaatc
     5161 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     5221 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     5281 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     5341 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5401 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5461 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     5521 tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
