LOCUS       sequence-423446-        8402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PIK3R4. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8402)
  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 8402)
  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_PIK3R4:1-1358
FEATURES             Location/Qualifiers
     source          1..8402
                     /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    5465..5586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5476..5495)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5701..5734)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5744..5791
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5823..6617
                     /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(5877..5896)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6487..6506
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6775..7363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6855..6874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7489..7537
                     /note="synthetic polyadenylation signal"
     CDS             complement(7561..7959)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7956..7965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7990..8347)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8004..8139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8156..8347
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8304..8324
                     /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 ccatgggaaa tcagcttgct
     1261 ggcattgctc cctcccagat cctttctgta gagagttatt tttcagatat tcatgacttt
     1321 gaatatgata aaagcctggg gagtactcgg ttttttaaag ttgctcgagc caagcaccga
     1381 gaaggcctgg tcgttgtgaa ggtttttgca attcaggatc ccacattgcc tttaaccagc
     1441 tataaacaag agctggagga actgaaaatc aggcttaatt ctgcacagaa ttgtctacct
     1501 ttccagaaag catcagaaaa agcatctgag aaagcagcta tgctctttag gcagtatgtg
     1561 cgagacaatc tctatgatcg catcagtacc cgtccattct tgaataacat tgagaagcgc
     1621 tggattgctt tccagatcct gacagctgtg gaccaagcac acaaatctgg agttcgtcat
     1681 ggggacatca agactgagaa tgtgatggtc accagttgga attgggttct tctaactgat
     1741 tttgccagtt ttaagcccac ttatcttcca gaagacaacc cggcagattt caattatttc
     1801 tttgacacat cacggaggag aacttgctat attgctcctg aacgttttgt tgatggtggg
     1861 atgtttgcca ctgagttaga atatatgaga gatccttcaa ctccgcttgt agacttaaat
     1921 agcaatcaga gaacaagagg agagttgaag agagcaatgg acatcttttc agcaggttgt
     1981 gtgatagctg agctttttac agaaggtgta ccattatttg atctctctca acttttggct
     2041 tatagaaatg gacatttttt ccctgaacaa gtgctaaata aaattgaaga tcacagtatc
     2101 agagaattgg taactcagat gattcaccgt gagccagata aacgtttaga ggcagaagat
     2161 tacttaaaac agcagcgtgg caatgccttt cctgaaatat tttacacttt tcttcagccc
     2221 tacatggccc agtttgccaa ggaaacgttt ctttctgcag atgagcgtat tctggttata
     2281 cggaaggatt tgggcaacat tattcacaat ctctgtggac atgatctgcc agaaaaagcc
     2341 gaaggagagc ctaaggaaaa tgggctggtt atcttggtat ctgttataac atcctgccta
     2401 cagaccctta aatactgtga ttccaaacta gctgctttgg aactgattct tcatttggct
     2461 ccaagattaa gtgttgaaat ccttttggat cgtattactc catatctttt gcatttcagc
     2521 aatgactctg ttcctagggt gagggctgaa gccttgagga cgttgaccaa agttcttgct
     2581 ctcgtcaaag aggttcctcg taatgatatc aatatttatc cggaatacat tctgccaggc
     2641 atagcccact tagcccaaga tgatgctact atcgttagac tagcctatgc tgaaaacata
     2701 gctctgctgg cagaaacagc tctgagattc ctggaattag tacagttaaa aaatcttaat
     2761 atggaaaatg accccaataa tgaagaaata gatgaggtta cacatccaaa tggaaattat
     2821 gacacagagc tccaagcctt acatgaaatg gtccagcaga aagttgttac tttgctaagt
     2881 gaccctgaaa atattgtaaa acaaaccttg atggaaaatg gaataacacg gctgtgtgta
     2941 ttctttggac gtcagaaagc caacgatgtt ttgttgtccc acatgattac tttcctaaat
     3001 gataagaatg attggcatct acgtggagca ttttttgata gtatagttgg tgttgctgcc
     3061 tatgttggct ggcaaagctc ctcaattctc aagcctctgc tgcaacaagg tcttagtgat
     3121 gctgaggaat ttgtcattgt gaaagctctt tatgccctta cttgtatgtg ccagttagga
     3181 ctgctacaaa aaccccatgt ttacgaattt gccagtgata ttgccccctt cctgtgtcat
     3241 cccaatttat ggatacgtta tggtgccgtg ggatttatca cagtggtagc tcgtcaaata
     3301 agtacagctg atgtctactg taaactgatg ccttatcttg acccatatat tacccaacca
     3361 ataatacaga ttgaaagaaa acttgttctg ctcagtgttt taaaggaacc agtaagtcgt
     3421 tctatatttg attatgcttt gaggtctaaa gatattacta gcttgttcag acatcttcac
     3481 atgcgtcaga agaaacgaaa tggttctctt cccgactgcc ctccgccaga ggatcctgcc
     3541 atagcacagc ttctgaagaa gttgctctca cagggaatga cagaggaaga ggaagacaaa
     3601 cttctggcac tgaaagactt catgatgaaa tctaataaag caaaggccaa tatagtggac
     3661 cagagccatc ttcatgatag tagtcagaaa ggtgtaattg acttggcagc tttaggcata
     3721 actgggagac aagttgatct tgttaaaacc aaacaagaac cagatgacaa acgggccaga
     3781 aaacatgtaa aacaagactc aaatgtaaat gaagaatgga aaagcatgtt tgggtcactg
     3841 gacccaccaa acatgccaca ggccctacct aaagggagtg atcaggaggt gattcagact
     3901 gggaaacctc ctcgttccga gtcctctgct ggcatttgtg tccctttgtc aacttcttca
     3961 caggttccag aagtgacaac tgtccaaaat aaaaaaccag taataccggt tttaagtagt
     4021 acaatcttac catccaccta tcagattcga attacaactt gtaaaactga acttcagcaa
     4081 ctcatccagc aaaagcggga gcagtgcaat gctgagagaa tagctaagca gatgatggaa
     4141 aatgctgaat gggagagtaa accaccacca cctggatggc gtcctaaagg gctgttagtt
     4201 gcccatcttc atgagcataa atctgctgtg aatcgaatta gagtctctga tgaacactca
     4261 ctttttgcaa catgttcaaa tgatggcaca gtgaaaatct ggaacagtca aaagatggag
     4321 gggaagacca ccactaccag atctattctt acatacagcc gaattggagg acgagtcaag
     4381 acgctcacat tctgccaagg ctcccactat ttagccatag catctgataa tggtgctgtc
     4441 cagcttcttg gaattgaggc ttctaagctg cccaagtctc ctaaaatcca tcctctacaa
     4501 agcagaattc tagatcagaa ggaggacggt tgtgttgtgg atatgcatca cttcaactct
     4561 ggagcacagt ctgttcttgc ctatgccact gtgaatggct ctctggttgg ctgggacctt
     4621 aggtcttcaa gcaatgcgtg gactttaaag catgatttaa agtcgggcct catcacttcc
     4681 tttgctgtgg acatccacca atgctggctc tgcattggta caagcagtgg taccatggct
     4741 tgttgggaca tgaggttcca gttgccaatt tcaagtcact gtcatccttc cagggctcga
     4801 atcagacgcc tctcaatgca ccctctgtat cagtcctggg tgattgcagc tgttcagggc
     4861 aacaacgaag tgtccatgtg ggacatggag actggtgaca gaagatttac tctctgggcc
     4921 agcagtgcac caccactttc tgaattacag ccttctcctc atagcgtcca tggtatctac
     4981 tgtagtcctg cagatggaaa tcctatccta ctaacagctg gctcagatat gaaaataagg
     5041 ttttgggact tggcttaccc agaaaggtcc tatgttgttg caggaagtac tagttcccca
     5101 tctgtgtcct actacaggaa aataattgaa ggcactgaag ttgtccagga aattcagaat
     5161 aagcagaaag taggaccaag tgatgacacc cctcgaaggg gcccagagtc cctgcccgtg
     5221 ggacatcatg acatcatcac tgatgtcgcc acattccaga ccacacaggg cttcatcgta
     5281 actgcttcta gagatgggat tgtgaaggtg tggaaaggct cgagcggtgg gagctccggt
     5341 gtgagcggct ggcggctgtt caagaagatt agctaagttt aaacggccgc gactctagac
     5401 tgcaggcatg caagctgatt tggctgctgc caccgctgag caaggccgct tcgagcagac
     5461 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     5521 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     5581 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     5641 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa gacttctaat
     5701 ataacttcgt atagcataca ttatacgaag ttatggttcc actgttgaca attaatcatc
     5761 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga
     5821 tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5881 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5941 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6001 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6061 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6121 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6181 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6241 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6301 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6361 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6421 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6481 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6541 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6601 ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca
     6661 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6721 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6781 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6841 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6901 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6961 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7021 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7081 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7141 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7201 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7261 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7321 tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt
     7381 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7441 attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca cacaaaaaac
     7501 caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga tggctagagt
     7561 ttagccctcc cacacataac cagagggcag caattcacga atcccaactg ccgtcggctg
     7621 tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa gcacctgtcg
     7681 gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga cgatacaagt
     7741 caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg cacaaggtcc
     7801 cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag agagagctgc
     7861 gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg ttgctctttc
     7921 aatgagggtg gattcttctt gagacaaagg cttggccatg gtggcgctag tgtcagaaga
     7981 atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     8041 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     8101 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     8161 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     8221 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     8281 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     8341 gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc atggtgctgc
     8401 gc
//
