LOCUS       sequence-425913-        5586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human NUP214 fragment M1 to T434. N-terminal
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  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 5586)
  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_NUP214:1-434
FEATURES             Location/Qualifiers
     source          1..5586
                     /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      1737..1746
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2649..2770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2660..2679)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2885..2918)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2928..2975
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3007..3801
                     /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(3061..3080)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3671..3690
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3959..4547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4039..4058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4673..4721
                     /note="synthetic polyadenylation signal"
     CDS             complement(4745..5143)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5140..5149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5174..5531)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5188..5323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5340..5531
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5488..5508
                     /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 tgggagacga
     1261 gatggatgcc atgattcccg agcgggagat gaaggatttt cagtttagag cgctaaagaa
     1321 ggtgagaatc tttgactccc ctgaggaatt gcccaaggaa cgctcgagtc tgcttgctgt
     1381 gtccaacaaa tatggtctgg tcttcgctgg tggagccagt ggcttgcaga tttttcctac
     1441 taaaaatctt cttattcaaa ataaacccgg agatgatccc aacaaaatag ttgataaagt
     1501 ccaaggcttg ctagttccta tgaaattccc aatccatcac ctggccttga gctgtgataa
     1561 cctcacactc tctgcgtgca tgatgtccag tgaatatggt tccattattg ctttttttga
     1621 tgttcgcaca ttctcaaatg aggctaaaca gcaaaaacgc ccatttgcct atcataagct
     1681 tttgaaagat gcaggaggca tggtgattga tatgaagtgg aaccccactg tcccctccat
     1741 ggtggcagtt tgtctggctg atggtagtat tgctgtcctg caagtcacgg aaacagtgaa
     1801 agtatgtgca actcttcctt ccacggtagc agtaacctct gtgtgctgga gccccaaagg
     1861 aaagcagctg gcagtgggaa aacagaatgg aactgtggtc cagtatcttc ctactttgca
     1921 ggaaaaaaaa gtcattcctt gtcctccgtt ttatgagtca gatcatcctg tcagagttct
     1981 ggatgtgctg tggattggta cctacgtctt cgccatagtg tatgctgctg cagatgggac
     2041 cctggaaacg tctccagatg tggtgatggc tctactaccg aaaaaagaag aaaagcaccc
     2101 agagatattt gtgaacttta tggagccctg ttatggcagc tgcacggaga gacagcatca
     2161 ttactacctc agttacattg aggaatggga tttagtgctg gcagcatctg cggcttcaac
     2221 agaagttagt atccttgctc gacaaagtga tcagattaat tgggaatctt ggctactgga
     2281 ggattctagt cgagctgaat tgcctgtgac agacaagagt gatgactcct tgcccatggg
     2341 agttgtcgta gactatacaa accaagtgga aatcaccatc agtgatgaaa agactcttcc
     2401 tcctgctcca gttctcatgt tactttcaac agatggtgtg ctttgtccat tttatatgat
     2461 taatcaaaat cctggggtta agtctctcat caaaacacca gagcgacttt cattagaagg
     2521 agagcgacag cccaagtcac caggaagtac ttaatctaga ggtcaagctt cgggggttta
     2581 aacctgcagg catgcaagct gatttggctg ctgccaccgc tgagcaaggc cgcttcgagc
     2641 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2701 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2761 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     2821 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcttaa ttaagacttc
     2881 taatataact tcgtatagca tacattatac gaagttatgg ttccactgtt gacaattaat
     2941 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg
     3001 cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3061 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3121 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3181 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3241 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3301 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3361 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3421 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3481 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3541 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3601 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3661 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3721 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3781 cttcttgacg agttcttctg attcgaaatg accgaccaag cgacgcccaa ccggtatcag
     3841 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3961 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4021 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4081 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4141 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4201 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4261 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4321 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4381 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4441 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4501 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga
     4561 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4621 tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaatc gtcacacaaa
     4681 aaaccaacac acagatgtaa tgaaaataaa gatattttat tgcggccatc gtgatggcta
     4741 gagtttagcc ctcccacaca taaccagagg gcagcaattc acgaatccca actgccgtcg
     4801 gctgtccatc actgtccttc actatggctt tgatcccagg atgcagatcg agaagcacct
     4861 gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc atttccgatc gcgacgatac
     4921 aagtcaggtt gccagctgcc gcagcagcag cagtgcccag caccacgagt tctgcacaag
     4981 gtcccccagt aaaatgatat acattgacac cagtgaagat gcggccgtcg ctagagagag
     5041 ctgcgctggc gacgctgtag tcttcagaga tggggatgct gttgattgta gccgttgctc
     5101 tttcaatgag ggtggattct tcttgagaca aaggcttggc catggtggcg ctagtgtcag
     5161 aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     5221 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     5281 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     5341 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     5401 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
     5461 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agctggttct
     5521 ttccgcctca gaaggtacct aaccaagttc ctctttcaga ggttatttca ggccatggtg
     5581 ctgcgc
//
