LOCUS       sequence-423417-        5438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human ARPC1A. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5438)
  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 5438)
  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_ARPC1A:1-370
FEATURES             Location/Qualifiers
     source          1..5438
                     /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    2501..2622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2512..2531)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2737..2770)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2780..2827
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2859..3653
                     /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(2913..2932)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3523..3542
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3811..4399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3891..3910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4525..4573
                     /note="synthetic polyadenylation signal"
     CDS             complement(4597..4995)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4992..5001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5026..5383)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5040..5175
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5192..5383
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5340..5360
                     /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 ccatgtcact gcatcagttt
     1261 ttactagagc caatcacctg tcatgcctgg aacagggatc gtactcagat tgccctcagt
     1321 cccaataatc acgaagtgca catctataag aagaacggga gccagtgggt gaaagctcat
     1381 gaactcaagg agcacaacgg acacatcaca ggtattgact gggctcccaa gagcgaccgc
     1441 attgtcactt gtggggcaga ccgcaatgcc tatgtctgga gtcagaaaga tggtgtttgg
     1501 aagccaaccc tggtgatcct gagaattaat cgcgcagcta cttttgtgaa gtggtccccc
     1561 ctagagaaca aatttgctgt gggaagtgga gcacgactca tttctgtttg ttactttgag
     1621 tctgaaaatg actggtgggt gagcaagcac attaaaaagc cgattcgctc cacagtcctc
     1681 agcttggatt ggcatcccaa caacgttttg ctggcagcag gatcatgtga cttcaaatgc
     1741 agagtgtttt ctgcctacat taaagaagtg gatgaaaagc cagccagcac gccctggggc
     1801 agcaagatgc cttttgggca gctgatgtca gagtttggtg gcagtggcac tggtggctgg
     1861 gtccacgggg taagcttctc tgccagtggg agccgcctgg cctgggtcag ccacgacagc
     1921 accgtgtctg ttgctgatgc ctcaaaaagt gtgcaggtct cgactctgaa gacagagttc
     1981 ctgccgctcc taagtgtgtc atttgtctca gagaacagcg tcgtggctgc tggccatgac
     2041 tgctgcccaa tgctctttaa ctacgatgac cgcggctgcc tgaccttcgt ctccaagtta
     2101 gatattccaa aacagagcat ccaacgcaac atgtctgcca tggaacgctt ccgcaacatg
     2161 gacaagagag ccacaactga ggaccgcaac acggccttgg agacgctgca ccagaatagc
     2221 atcactcaag tctctattta tgaggtggac aagcaagatt gtcgcaaatt ttgcactact
     2281 ggcatcgatg gagccatgac aatttgggat ttcaagaccc tcgagtcttc catccagggc
     2341 ctccggataa tgggctcgag cggtgggagc tccggtgtga gcggctggcg gctgttcaag
     2401 aagattagct aagtttaaac ggccgcgact ctagactgca ggcatgcaag ctgatttggc
     2461 tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat tgatgagttt
     2521 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2581 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2641 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     2701 tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag catacattat
     2761 acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg gcatagtata
     2821 atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc
     2881 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2941 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3001 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3061 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3121 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3181 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3241 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3301 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3361 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3421 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3481 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3541 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3601 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaaa
     3661 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     3721 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3781 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3841 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3901 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3961 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4021 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4081 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4141 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4201 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4261 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4321 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4381 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4441 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4501 gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt aatgaaaata
     4561 aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca cataaccaga
     4621 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     4681 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     4741 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     4801 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     4861 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     4921 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     4981 caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag
     5041 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     5101 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     5161 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     5221 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     5281 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     5341 ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac ctaaccaagt
     5401 tcctctttca gaggttattt caggccatgg tgctgcgc
//
