LOCUS       sequence-423418-        5444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human ARPC1B. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5444)
  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 5444)
  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_ARPC1B:1-372
FEATURES             Location/Qualifiers
     source          1..5444
                     /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    2507..2628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2518..2537)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2743..2776)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2786..2833
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2865..3659
                     /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(2919..2938)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3529..3548
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3817..4405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3897..3916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4531..4579
                     /note="synthetic polyadenylation signal"
     CDS             complement(4603..5001)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4998..5007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5032..5389)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5046..5181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5198..5389
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5346..5366
                     /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 ccatggccta ccacagcttc
     1261 ctggtggagc ccatcagctg ccacgcctgg aacaaggacc gcacccagat tgccatctgc
     1321 cccaacaacc atgaggtgca tatctatgaa aagagcggtg ccaaatggac caaggtgcac
     1381 gagctcaagg agcacaacgg gcaggtgaca ggcatcgact gggcccccga gagtaaccgt
     1441 attgtgacct gcggcacaga ccgcaacgcc tacgtgtgga cgctgaaggg ccgcacatgg
     1501 aagcccacgc tggtcatcct gcggatcaac cgggctgccc gctgcgtgcg ctgggccccc
     1561 aacgagaaca agtttgctgt gggcagcggc tctcgtgtga tctccatctg ttatttcgag
     1621 caggagaatg actggtgggt ttgcaagcac atcaagaagc ccatccgctc caccgtcctc
     1681 agcctggact ggcaccccaa caatgtgctg ctggctgccg gctcctgtga cttcaagtgt
     1741 cggatctttt cagcctacat caaggaggtg gaggaacggc cggcacccac cccgtggggc
     1801 tccaagatgc cctttgggga actgatgttc gaatccagca gtagctgcgg ctgggtacat
     1861 ggcgtctgtt tctcagccag cgggagccgc gtggcctggg taagccacga cagcaccgtc
     1921 tgcctggctg atgccgacaa gaagatggcc gtcgcgactc tggcctctga aacactacca
     1981 ctgctggcgc tgaccttcat cacagacaac agcctggtgg cagcgggcca cgactgcttc
     2041 ccggtgctgt tcacctatga cgccgccgcg gggatgctga gcttcggcgg gcggctggac
     2101 gttcctaagc agagctcgca gcgtggcttg acggcccgcg agcgcttcca gaacctggac
     2161 aagaaggcga gctccgaggg tggcacggct gcgggcgcgg gcctagactc gctgcacaag
     2221 aacagcgtca gccagatctc ggtgctcagc ggcggcaagg ccaagtgctc gcagttctgc
     2281 accactggca tggatggcgg catgagtatc tgggatgtga agagcttgga gtcagccttg
     2341 aaggacctca agatcaaagg ctcgagcggt gggagctccg gtgtgagcgg ctggcggctg
     2401 ttcaagaaga ttagctaagt ttaaacggcc gcgactctag actgcaggca tgcaagctga
     2461 tttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag atacattgat
     2521 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2581 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2641 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     2701 aacctctaca aatgtggtaa aatcttaatt aagacttcta atataacttc gtatagcata
     2761 cattatacga agttatggtt ccactgttga caattaatca tcggcatagt atatcggcat
     2821 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg
     2881 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2941 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3001 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3061 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3121 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3181 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3241 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3301 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3361 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3421 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3481 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3541 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3601 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat
     3661 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     3721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4381 tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4501 cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac agatgtaatg
     4561 aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct cccacacata
     4621 accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac
     4681 tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc cgcaggggct
     4741 caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc
     4801 agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa aatgatatac
     4861 attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc
     4921 ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc
     4981 ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa
     5041 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     5101 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     5161 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     5221 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     5281 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5341 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
     5401 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
