LOCUS       sequence-425918-        8358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PIK3R4. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8358)
  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 8358)
  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_PIK3R4:1-1358
FEATURES             Location/Qualifiers
     source          1..8358
                     /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    5421..5542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5432..5451)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5657..5690)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5700..5747
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5779..6573
                     /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(5833..5852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6443..6462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6731..7319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6811..6830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7445..7493
                     /note="synthetic polyadenylation signal"
     CDS             complement(7517..7915)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7912..7921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7946..8303)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7960..8095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8112..8303
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8260..8280
                     /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 tgggaaatca
     1261 gcttgctggc attgctccct cccagatcct ttctgtagag agttattttt cagatattca
     1321 tgactttgaa tatgataaaa gcctggggag tactcggttt tttaaagttg ctcgagccaa
     1381 gcaccgagaa ggcctggtcg ttgtgaaggt ttttgcaatt caggatccca cattgccttt
     1441 aaccagctat aaacaagagc tggaggaact gaaaatcagg cttaattctg cacagaattg
     1501 tctacctttc cagaaagcat cagaaaaagc atctgagaaa gcagctatgc tctttaggca
     1561 gtatgtgcga gacaatctct atgatcgcat cagtacccgt ccattcttga ataacattga
     1621 gaagcgctgg attgctttcc agatcctgac agctgtggac caagcacaca aatctggagt
     1681 tcgtcatggg gacatcaaga ctgagaatgt gatggtcacc agttggaatt gggttcttct
     1741 aactgatttt gccagtttta agcccactta tcttccagaa gacaacccgg cagatttcaa
     1801 ttatttcttt gacacatcac ggaggagaac ttgctatatt gctcctgaac gttttgttga
     1861 tggtgggatg tttgccactg agttagaata tatgagagat ccttcaactc cgcttgtaga
     1921 cttaaatagc aatcagagaa caagaggaga gttgaagaga gcaatggaca tcttttcagc
     1981 aggttgtgtg atagctgagc tttttacaga aggtgtacca ttatttgatc tctctcaact
     2041 tttggcttat agaaatggac attttttccc tgaacaagtg ctaaataaaa ttgaagatca
     2101 cagtatcaga gaattggtaa ctcagatgat tcaccgtgag ccagataaac gtttagaggc
     2161 agaagattac ttaaaacagc agcgtggcaa tgcctttcct gaaatatttt acacttttct
     2221 tcagccctac atggcccagt ttgccaagga aacgtttctt tctgcagatg agcgtattct
     2281 ggttatacgg aaggatttgg gcaacattat tcacaatctc tgtggacatg atctgccaga
     2341 aaaagccgaa ggagagccta aggaaaatgg gctggttatc ttggtatctg ttataacatc
     2401 ctgcctacag acccttaaat actgtgattc caaactagct gctttggaac tgattcttca
     2461 tttggctcca agattaagtg ttgaaatcct tttggatcgt attactccat atcttttgca
     2521 tttcagcaat gactctgttc ctagggtgag ggctgaagcc ttgaggacgt tgaccaaagt
     2581 tcttgctctc gtcaaagagg ttcctcgtaa tgatatcaat atttatccgg aatacattct
     2641 gccaggcata gcccacttag cccaagatga tgctactatc gttagactag cctatgctga
     2701 aaacatagct ctgctggcag aaacagctct gagattcctg gaattagtac agttaaaaaa
     2761 tcttaatatg gaaaatgacc ccaataatga agaaatagat gaggttacac atccaaatgg
     2821 aaattatgac acagagctcc aagccttaca tgaaatggtc cagcagaaag ttgttacttt
     2881 gctaagtgac cctgaaaata ttgtaaaaca aaccttgatg gaaaatggaa taacacggct
     2941 gtgtgtattc tttggacgtc agaaagccaa cgatgttttg ttgtcccaca tgattacttt
     3001 cctaaatgat aagaatgatt ggcatctacg tggagcattt tttgatagta tagttggtgt
     3061 tgctgcctat gttggctggc aaagctcctc aattctcaag cctctgctgc aacaaggtct
     3121 tagtgatgct gaggaatttg tcattgtgaa agctctttat gcccttactt gtatgtgcca
     3181 gttaggactg ctacaaaaac cccatgttta cgaatttgcc agtgatattg cccccttcct
     3241 gtgtcatccc aatttatgga tacgttatgg tgccgtggga tttatcacag tggtagctcg
     3301 tcaaataagt acagctgatg tctactgtaa actgatgcct tatcttgacc catatattac
     3361 ccaaccaata atacagattg aaagaaaact tgttctgctc agtgttttaa aggaaccagt
     3421 aagtcgttct atatttgatt atgctttgag gtctaaagat attactagct tgttcagaca
     3481 tcttcacatg cgtcagaaga aacgaaatgg ttctcttccc gactgccctc cgccagagga
     3541 tcctgccata gcacagcttc tgaagaagtt gctctcacag ggaatgacag aggaagagga
     3601 agacaaactt ctggcactga aagacttcat gatgaaatct aataaagcaa aggccaatat
     3661 agtggaccag agccatcttc atgatagtag tcagaaaggt gtaattgact tggcagcttt
     3721 aggcataact gggagacaag ttgatcttgt taaaaccaaa caagaaccag atgacaaacg
     3781 ggccagaaaa catgtaaaac aagactcaaa tgtaaatgaa gaatggaaaa gcatgtttgg
     3841 gtcactggac ccaccaaaca tgccacaggc cctacctaaa gggagtgatc aggaggtgat
     3901 tcagactggg aaacctcctc gttccgagtc ctctgctggc atttgtgtcc ctttgtcaac
     3961 ttcttcacag gttccagaag tgacaactgt ccaaaataaa aaaccagtaa taccggtttt
     4021 aagtagtaca atcttaccat ccacctatca gattcgaatt acaacttgta aaactgaact
     4081 tcagcaactc atccagcaaa agcgggagca gtgcaatgct gagagaatag ctaagcagat
     4141 gatggaaaat gctgaatggg agagtaaacc accaccacct ggatggcgtc ctaaagggct
     4201 gttagttgcc catcttcatg agcataaatc tgctgtgaat cgaattagag tctctgatga
     4261 acactcactt tttgcaacat gttcaaatga tggcacagtg aaaatctgga acagtcaaaa
     4321 gatggagggg aagaccacca ctaccagatc tattcttaca tacagccgaa ttggaggacg
     4381 agtcaagacg ctcacattct gccaaggctc ccactattta gccatagcat ctgataatgg
     4441 tgctgtccag cttcttggaa ttgaggcttc taagctgccc aagtctccta aaatccatcc
     4501 tctacaaagc agaattctag atcagaagga ggacggttgt gttgtggata tgcatcactt
     4561 caactctgga gcacagtctg ttcttgccta tgccactgtg aatggctctc tggttggctg
     4621 ggaccttagg tcttcaagca atgcgtggac tttaaagcat gatttaaagt cgggcctcat
     4681 cacttccttt gctgtggaca tccaccaatg ctggctctgc attggtacaa gcagtggtac
     4741 catggcttgt tgggacatga ggttccagtt gccaatttca agtcactgtc atccttccag
     4801 ggctcgaatc agacgcctct caatgcaccc tctgtatcag tcctgggtga ttgcagctgt
     4861 tcagggcaac aacgaagtgt ccatgtggga catggagact ggtgacagaa gatttactct
     4921 ctgggccagc agtgcaccac cactttctga attacagcct tctcctcata gcgtccatgg
     4981 tatctactgt agtcctgcag atggaaatcc tatcctacta acagctggct cagatatgaa
     5041 aataaggttt tgggacttgg cttacccaga aaggtcctat gttgttgcag gaagtactag
     5101 ttccccatct gtgtcctact acaggaaaat aattgaaggc actgaagttg tccaggaaat
     5161 tcagaataag cagaaagtag gaccaagtga tgacacccct cgaaggggcc cagagtccct
     5221 gcccgtggga catcatgaca tcatcactga tgtcgccaca ttccagacca cacagggctt
     5281 catcgtaact gcttctagag atgggattgt gaaggtgtgg aaataatcta gaggtcaagc
     5341 ttcgggggtt taaacctgca ggcatgcaag ctgatttggc tgctgccacc gctgagcaag
     5401 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     5461 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     5521 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     5581 gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatctt
     5641 aattaagact tctaatataa cttcgtatag catacattat acgaagttat ggttccactg
     5701 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     5761 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5821 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5881 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5941 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6001 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6061 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6121 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6181 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6241 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6301 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6361 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6421 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6481 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6541 gccttctatc gccttcttga cgagttcttc tgattcgaaa tgaccgacca agcgacgccc
     6601 aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6901 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7141 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag
     7321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa
     7441 tcgtcacaca aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca
     7501 tcgtgatggc tagagtttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc
     7561 caactgccgt cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat
     7621 cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga
     7681 tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga
     7741 gttctgcaca aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt
     7801 cgctagagag agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg
     7861 tagccgttgc tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatggtgg
     7921 cgctagtgtc agaagaatcg agctttttgc aaaagcctag gcctccaaaa aagcctcctc
     7981 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     8041 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     8101 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     8161 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     8221 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     8281 acagctggtt ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt
     8341 caggccatgg tgctgcgc
//
