LOCUS       sequence-423448-        5228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human PLAA fragment R9 to A307. C-terminal 
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5228)
  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 5228)
  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:9-307
FEATURES             Location/Qualifiers
     source          1..5228
                     /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    2291..2412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2302..2321)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2527..2560)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2570..2617
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2649..3443
                     /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(2703..2722)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3313..3332
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3601..4189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3681..3700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4315..4363
                     /note="synthetic polyadenylation signal"
     CDS             complement(4387..4785)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4782..4791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4816..5173)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4830..4965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4982..5173
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5130..5150
                     /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 ccatgcggct gagctgctcg
     1261 ctccggggcc acgagctgga cgtacggggc ctggtgtgct gcgcctatcc gccgggagcc
     1321 tttgtgtccg tgtcccgaga ccgcaccacc cgcctctggg ccccagacag tccaaacagg
     1381 agctttacag aaatgcactg tatgagtggc cattccaatt ttgtatcttg tgtatgcatc
     1441 ataccctcaa gtgacatcta ccctcatggc ctaattgcca ccggtggaaa tgaccacaat
     1501 atatgcattt tctcactgga cagtccaatg ccactttata ttctaaaagg ccacaaaaat
     1561 actgtttgta gtctatcatc tggaaaattt gggacattac ttagtggttc atgggacacc
     1621 actgctaaag tctggctgaa tgacaagtgc atgatgacct tgcagggtca tacagctgca
     1681 gtgtgggcgg taaagatctt acctgaacag ggcttaatgt tgactggatc agcagacaag
     1741 actgttaaac tgtggaaggc tggaagatgt gagaggactt tttcagggca tgaagactgt
     1801 gtaagaggtt tggcaatttt gagtgaaaca gaatttcttt cctgtgcaaa tgatgctagt
     1861 attagaaggt ggcaaatcac tggcgagtgt cttgaagtat attatggaca tacaaattat
     1921 atttatagca tatccgtttt tccaaattgt agagactttg tgacaacagc agaggacaga
     1981 tctctgagaa tctggaaaca tggggaatgt gctcaaacta tccgacttcc agctcagtct
     2041 atatggtgct gctgtgtgct cgacaatggt gacattgtgg ttggtgcgag tgatggcatt
     2101 attagagtgt ttacagaatc agaagatcga acagcaagtg ctggctcgag cggtgggagc
     2161 tccggtgtga gcggctggcg gctgttcaag aagattagct aagtttaaac ggccgcgact
     2221 ctagactgca ggcatgcaag ctgatttggc tgctgccacc gctgagcaag gccgcttcga
     2281 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2341 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2401 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     2461 tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatctt aattaagact
     2521 tctaatataa cttcgtatag catacattat acgaagttat ggttccactg ttgacaatta
     2581 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga
     2641 ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2701 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2761 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2821 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     2881 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     2941 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3001 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3061 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3121 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3181 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3241 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3301 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3361 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3421 gccttcttga cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc
     3481 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3541 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3601 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3661 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3721 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3781 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3841 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3901 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3961 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4021 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4081 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4141 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     4201 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4261 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca
     4321 aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc
     4381 tagagtttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt
     4441 cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac
     4501 ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat
     4561 acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga gttctgcaca
     4621 aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag
     4681 agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc
     4741 tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatggtgg cgctagtgtc
     4801 agaagaatcg agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     4861 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     4921 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     4981 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     5041 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     5101 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     5161 ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt caggccatgg
     5221 tgctgcgc
//
