LOCUS       sequence-425919-        6669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PLAA. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6669)
  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 6669)
  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_PLAA:1-795
FEATURES             Location/Qualifiers
     source          1..6669
                     /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    3732..3853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3743..3762)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3968..4001)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4011..4058
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4090..4884
                     /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(4144..4163)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4754..4773
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5042..5630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5122..5141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5756..5804
                     /note="synthetic polyadenylation signal"
     CDS             complement(5828..6226)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6223..6232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6257..6614)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6271..6406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6423..6614
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6571..6591
                     /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 tgacgagcgg
     1261 cgcaaccagg taccggctga gctgctcgct ccggggccac gagctggacg tacggggcct
     1321 ggtgtgctgc gcctatccgc cgggagcctt tgtgtccgtg tcccgagacc gcaccacccg
     1381 cctctgggcc ccagacagtc caaacaggag ctttacagaa atgcactgta tgagtggcca
     1441 ttccaatttt gtatcttgtg tatgcatcat accctcaagt gacatctacc ctcatggcct
     1501 aattgccacc ggtggaaatg accacaatat atgcattttc tcactggaca gtccaatgcc
     1561 actttatatt ctaaaaggcc acaaaaatac tgtttgtagt ctatcatctg gaaaatttgg
     1621 gacattactt agtggttcat gggacaccac tgctaaagtc tggctgaatg acaagtgcat
     1681 gatgaccttg cagggtcata cagctgcagt gtgggcggta aagatcttac ctgaacaggg
     1741 cttaatgttg actggatcag cagacaagac tgttaaactg tggaaggctg gaagatgtga
     1801 gaggactttt tcagggcatg aagactgtgt aagaggtttg gcaattttga gtgaaacaga
     1861 atttctttcc tgtgcaaatg atgctagtat tagaaggtgg caaatcactg gcgagtgtct
     1921 tgaagtatat tatggacata caaattatat ttatagcata tccgtttttc caaattgtag
     1981 agactttgtg acaacagcag aggacagatc tctgagaatc tggaaacatg gggaatgtgc
     2041 tcaaactatc cgacttccag ctcagtctat atggtgctgc tgtgtgctcg acaatggtga
     2101 cattgtggtt ggtgcgagtg atggcattat tagagtgttt acagaatcag aagatcgaac
     2161 agcaagtgct gaagaaatca aggcttttga aaaagaactg tctcacgcaa ccattgattc
     2221 taaaactggc gatttagggg acatcaatgc tgagcagctt cctgggaggg aacatcttaa
     2281 tgaacctggt actagagaag gacagactcg tctaatcaga gatggggaga aagtcgaagc
     2341 ctatcagtgg agtgttagtg aagggaggtg gataaaaatt ggtgatgttg ttggctcatc
     2401 tggtgctaat cagcaaacat ctggaaaagt tttatatgaa gggaaagaat ttgattatgt
     2461 tttctcaatt gatgtcaatg aaggtggacc atcatataaa ttgccatata ataccagtga
     2521 tgacccttgg ttaactgcat acaacttctt acagaagaat gatttgaatc ctatgtttct
     2581 ggatcaagta gctaaattta ttattgataa cacaaaaggt caaatgttgg gacttgggaa
     2641 tcccagcttt tcagatccat ttacaggtgg tggtcggtat gttccgggct cttcgggatc
     2701 ttctaacaca ctacccacag cagatccttt tacaggtgct ggtcgttatg taccaggttc
     2761 tgcaagtatg ggaactacca tggccggagt tgatccattt acagggaata gtgcctaccg
     2821 atcagctgca tctaaaacaa tgaatattta tttccctaaa aaagaggctg tcacatttga
     2881 ccaagcaaac cctacacaaa tattaggtaa actgaaggaa cttaatggaa ctgcacctga
     2941 agagaagaag ttaactgagg atgacttgat acttcttgag aagatactgt ctctaatatg
     3001 taatagttct tcagaaaaac ccacagtcca gcaacttcag attttgtgga aagctattaa
     3061 ctgtcctgaa gatattgtct ttcctgcact tgacattctt cggttgtcaa ttaaacaccc
     3121 cagtgtgaat gagaacttct gcaatgaaaa ggaaggggct cagttcagca gtcatcttat
     3181 caatcttctg aaccctaaag gaaagccagc aaaccagctg cttgctctca ggactttttg
     3241 caattgtttt gttggccagg caggacaaaa actcatgatg tcccagaggg aatcactgat
     3301 gtcccatgca atagaactga aatcagggag caataagaac attcacattg ctctggctac
     3361 attggccctg aactattctg tttgttttca taaagaccat aacattgaag ggaaagccca
     3421 atgtttgtca ctaattagca caatcttgga agtagtacaa gacctagaag ccacttttag
     3481 acttcttgtg gctcttggaa cacttatcag tgatgattca aatgctgtac aattagccaa
     3541 gtctttaggt gttgattctc aaataaaaaa gtattcctca gtatcagaac cagctaaagt
     3601 aagtgaatgc tgtagattta tcctaaattt gctgtaatct agaggtcaag cttcgggggt
     3661 ttaaacctgc aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg
     3721 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3781 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3841 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     3901 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac
     3961 ttctaatata acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt
     4021 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     4081 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4141 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4201 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4261 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4321 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4381 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4441 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4501 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4561 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4621 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4681 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4741 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4801 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4861 cgccttcttg acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat
     4921 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4981 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5041 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5101 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5161 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5221 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5281 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5341 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5401 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5461 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5521 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5581 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt
     5641 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5701 tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac
     5761 aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg
     5821 ctagagttta gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg
     5881 tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca
     5941 cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga
     6001 tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac
     6061 aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga
     6121 gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg
     6181 ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt
     6241 cagaagaatc gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     6301 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     6361 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     6421 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     6481 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     6541 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt
     6601 tctttccgcc tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg
     6661 gtgctgcgc
//
