LOCUS       sequence-423441-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human NUP214 fragment M1 to T43. C-terminal 
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  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 5630)
  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_NUP214:1-434
FEATURES             Location/Qualifiers
     source          1..5630
                     /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"
     regulatory      1730..1739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2693..2814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2704..2723)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2929..2962)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2972..3019
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3051..3845
                     /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(3105..3124)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3715..3734
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4003..4591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4083..4102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4717..4765
                     /note="synthetic polyadenylation signal"
     CDS             complement(4789..5187)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5184..5193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5218..5575)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5232..5367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5384..5575
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5532..5552
                     /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 ccatgggaga cgagatggat
     1261 gccatgattc ccgagcggga gatgaaggat tttcagttta gagcgctaaa gaaggtgaga
     1321 atctttgact cccctgagga attgcccaag gaacgctcga gtctgcttgc tgtgtccaac
     1381 aaatatggtc tggtcttcgc tggtggagcc agtggcttgc agatttttcc tactaaaaat
     1441 cttcttattc aaaataaacc cggagatgat cccaacaaaa tagttgataa agtccaaggc
     1501 ttgctagttc ctatgaaatt cccaatccat cacctggcct tgagctgtga taacctcaca
     1561 ctctctgcgt gcatgatgtc cagtgaatat ggttccatta ttgctttttt tgatgttcgc
     1621 acattctcaa atgaggctaa acagcaaaaa cgcccatttg cctatcataa gcttttgaaa
     1681 gatgcaggag gcatggtgat tgatatgaag tggaacccca ctgtcccctc catggtggca
     1741 gtttgtctgg ctgatggtag tattgctgtc ctgcaagtca cggaaacagt gaaagtatgt
     1801 gcaactcttc cttccacggt agcagtaacc tctgtgtgct ggagccccaa aggaaagcag
     1861 ctggcagtgg gaaaacagaa tggaactgtg gtccagtatc ttcctacttt gcaggaaaaa
     1921 aaagtcattc cttgtcctcc gttttatgag tcagatcatc ctgtcagagt tctggatgtg
     1981 ctgtggattg gtacctacgt cttcgccata gtgtatgctg ctgcagatgg gaccctggaa
     2041 acgtctccag atgtggtgat ggctctacta ccgaaaaaag aagaaaagca cccagagata
     2101 tttgtgaact ttatggagcc ctgttatggc agctgcacgg agagacagca tcattactac
     2161 ctcagttaca ttgaggaatg ggatttagtg ctggcagcat ctgcggcttc aacagaagtt
     2221 agtatccttg ctcgacaaag tgatcagatt aattgggaat cttggctact ggaggattct
     2281 agtcgagctg aattgcctgt gacagacaag agtgatgact ccttgcccat gggagttgtc
     2341 gtagactata caaaccaagt ggaaatcacc atcagtgatg aaaagactct tcctcctgct
     2401 ccagttctca tgttactttc aacagatggt gtgctttgtc cattttatat gattaatcaa
     2461 aatcctgggg ttaagtctct catcaaaaca ccagagcgac tttcattaga aggagagcga
     2521 cagcccaagt caccaggaag tactggctcg agcggtggga gctccggtgt gagcggctgg
     2581 cggctgttca agaagattag ctaagtttaa acggccgcga ctctagactg caggcatgca
     2641 agctgatttg gctgctgcca ccgctgagca aggccgcttc gagcagacat gataagatac
     2701 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2761 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2821 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     2881 aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga cttctaatat aacttcgtat
     2941 agcatacatt atacgaagtt atggttccac tgttgacaat taatcatcgg catagtatat
     3001 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     3061 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3121 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3181 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3241 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3301 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3361 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3421 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3481 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3541 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3601 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3661 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3721 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3781 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3841 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     3901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4441 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4561 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     4621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4681 gatcttcacc tagatccttt tatagtccgg aatcgtcaca caaaaaacca acacacagat
     4741 gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagttt agccctccca
     4801 cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc
     4861 cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca
     4921 ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgccagc
     4981 tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg
     5041 atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct
     5101 gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga
     5161 ttcttcttga gacaaaggct tggccatggt ggcgctagtg tcagaagaat cgagcttttt
     5221 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     5281 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     5341 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     5401 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     5461 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5521 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcagaaggt
     5581 acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
