LOCUS       sequence-423447-        6713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PLAA. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6713)
  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 6713)
  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_PLAA:1-795
FEATURES             Location/Qualifiers
     source          1..6713
                     /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    3776..3897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3787..3806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4012..4045)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4055..4102
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4134..4928
                     /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(4188..4207)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4798..4817
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5086..5674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5166..5185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5800..5848
                     /note="synthetic polyadenylation signal"
     CDS             complement(5872..6270)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6267..6276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6301..6658)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6315..6450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6467..6658
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6615..6635
                     /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 ccatgacgag cggcgcaacc
     1261 aggtaccggc tgagctgctc gctccggggc cacgagctgg acgtacgggg cctggtgtgc
     1321 tgcgcctatc cgccgggagc ctttgtgtcc gtgtcccgag accgcaccac ccgcctctgg
     1381 gccccagaca gtccaaacag gagctttaca gaaatgcact gtatgagtgg ccattccaat
     1441 tttgtatctt gtgtatgcat cataccctca agtgacatct accctcatgg cctaattgcc
     1501 accggtggaa atgaccacaa tatatgcatt ttctcactgg acagtccaat gccactttat
     1561 attctaaaag gccacaaaaa tactgtttgt agtctatcat ctggaaaatt tgggacatta
     1621 cttagtggtt catgggacac cactgctaaa gtctggctga atgacaagtg catgatgacc
     1681 ttgcagggtc atacagctgc agtgtgggcg gtaaagatct tacctgaaca gggcttaatg
     1741 ttgactggat cagcagacaa gactgttaaa ctgtggaagg ctggaagatg tgagaggact
     1801 ttttcagggc atgaagactg tgtaagaggt ttggcaattt tgagtgaaac agaatttctt
     1861 tcctgtgcaa atgatgctag tattagaagg tggcaaatca ctggcgagtg tcttgaagta
     1921 tattatggac atacaaatta tatttatagc atatccgttt ttccaaattg tagagacttt
     1981 gtgacaacag cagaggacag atctctgaga atctggaaac atggggaatg tgctcaaact
     2041 atccgacttc cagctcagtc tatatggtgc tgctgtgtgc tcgacaatgg tgacattgtg
     2101 gttggtgcga gtgatggcat tattagagtg tttacagaat cagaagatcg aacagcaagt
     2161 gctgaagaaa tcaaggcttt tgaaaaagaa ctgtctcacg caaccattga ttctaaaact
     2221 ggcgatttag gggacatcaa tgctgagcag cttcctggga gggaacatct taatgaacct
     2281 ggtactagag aaggacagac tcgtctaatc agagatgggg agaaagtcga agcctatcag
     2341 tggagtgtta gtgaagggag gtggataaaa attggtgatg ttgttggctc atctggtgct
     2401 aatcagcaaa catctggaaa agttttatat gaagggaaag aatttgatta tgttttctca
     2461 attgatgtca atgaaggtgg accatcatat aaattgccat ataataccag tgatgaccct
     2521 tggttaactg catacaactt cttacagaag aatgatttga atcctatgtt tctggatcaa
     2581 gtagctaaat ttattattga taacacaaaa ggtcaaatgt tgggacttgg gaatcccagc
     2641 ttttcagatc catttacagg tggtggtcgg tatgttccgg gctcttcggg atcttctaac
     2701 acactaccca cagcagatcc ttttacaggt gctggtcgtt atgtaccagg ttctgcaagt
     2761 atgggaacta ccatggccgg agttgatcca tttacaggga atagtgccta ccgatcagct
     2821 gcatctaaaa caatgaatat ttatttccct aaaaaagagg ctgtcacatt tgaccaagca
     2881 aaccctacac aaatattagg taaactgaag gaacttaatg gaactgcacc tgaagagaag
     2941 aagttaactg aggatgactt gatacttctt gagaagatac tgtctctaat atgtaatagt
     3001 tcttcagaaa aacccacagt ccagcaactt cagattttgt ggaaagctat taactgtcct
     3061 gaagatattg tctttcctgc acttgacatt cttcggttgt caattaaaca ccccagtgtg
     3121 aatgagaact tctgcaatga aaaggaaggg gctcagttca gcagtcatct tatcaatctt
     3181 ctgaacccta aaggaaagcc agcaaaccag ctgcttgctc tcaggacttt ttgcaattgt
     3241 tttgttggcc aggcaggaca aaaactcatg atgtcccaga gggaatcact gatgtcccat
     3301 gcaatagaac tgaaatcagg gagcaataag aacattcaca ttgctctggc tacattggcc
     3361 ctgaactatt ctgtttgttt tcataaagac cataacattg aagggaaagc ccaatgtttg
     3421 tcactaatta gcacaatctt ggaagtagta caagacctag aagccacttt tagacttctt
     3481 gtggctcttg gaacacttat cagtgatgat tcaaatgctg tacaattagc caagtcttta
     3541 ggtgttgatt ctcaaataaa aaagtattcc tcagtatcag aaccagctaa agtaagtgaa
     3601 tgctgtagat ttatcctaaa tttgctgggc tcgagcggtg ggagctccgg tgtgagcggc
     3661 tggcggctgt tcaagaagat tagctaagtt taaacggccg cgactctaga ctgcaggcat
     3721 gcaagctgat ttggctgctg ccaccgctga gcaaggccgc ttcgagcaga catgataaga
     3781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3901 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta
     3961 agcaagtaaa acctctacaa atgtggtaaa atcttaatta agacttctaa tataacttcg
     4021 tatagcatac attatacgaa gttatggttc cactgttgac aattaatcat cggcatagta
     4081 tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgattg
     4141 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4201 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4261 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4321 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4381 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4441 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4501 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4561 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4621 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4681 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4741 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4801 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4861 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4921 tcttctgatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc
     4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5521 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5641 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5761 aaggatcttc acctagatcc ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca
     5821 gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag tttagccctc
     5881 ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct gtccatcact
     5941 gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc
     6001 gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag tcaggttgcc
     6061 agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc ccccagtaaa
     6121 atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg cgctggcgac
     6181 gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt caatgagggt
     6241 ggattcttct tgagacaaag gcttggccat ggtggcgcta gtgtcagaag aatcgagctt
     6301 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     6361 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     6421 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     6481 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     6541 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     6601 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagaa
     6661 ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg cgc
//
