LOCUS       sequence-426023-        5739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human NLE1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5739)
  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 5739)
  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_NLE1:1-485
FEATURES             Location/Qualifiers
     source          1..5739
                     /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    2802..2923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2813..2832)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3038..3071)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3081..3128
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3160..3954
                     /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(3214..3233)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3824..3843
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4112..4700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4192..4211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4826..4874
                     /note="synthetic polyadenylation signal"
     CDS             complement(4898..5296)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5293..5302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5327..5684)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5341..5476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5493..5684
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5641..5661
                     /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 tggcggcagc
     1261 agtggcggac gaggcggtgg cgcgcgatgt gcagcggttg ctagtgcagt tccaggatga
     1321 gggcgggcag ctgctgggtt ccccgttcga cgtgcccgtg gacatcaccc cggacaggct
     1381 gcagctcgtg tgcaacgcgc tactggccca ggaggatccc ctgccactgg ctttctttgt
     1441 ccacgatgct gagatcgtct cctcactggg gaagacgttg gagtcccagg cagtggagac
     1501 agagaaggtc ctagacatca tctaccagcc acaggctatc ttcagagtcc gggctgtgac
     1561 tcgctgcacc agctccttgg agggtcacag tgaggcagtc atttctgtgg ccttcagccc
     1621 tacgggaaag tacctggcca gtggctctgg agacaccacc gtgcgcttct gggatctcag
     1681 cacagagaca ccacatttca catgcaaggg acacagacac tgggtcctta gtatatcctg
     1741 gtctccagat ggcaagaagc tggcctcagg ctgcaagaat ggccagattc tcctctggga
     1801 cccaagcaca gggaagcagg tgggcaggac cctcgctggc cacagcaagt ggatcacagg
     1861 cctgagctgg gagcccctcc atgcgaaccc tgagtgccgc tatgtggcca gcagctccaa
     1921 ggatggcagt gtgcggatct gggacacaac tgcaggccgc tgtgagcgca tcctcaccgg
     1981 gcacacccag tcggtcacct gtctccggtg gggaggggac gggcttctct actctgcctc
     2041 ccaggaccgc accatcaaag tctggagagc tcatgacggt gtgctgtgcc ggactctgca
     2101 aggccacggc cactgggtga acaccatggc cctcagcact gactatgccc tgcgcactgg
     2161 ggcctttgaa cctgctgagg cctcagttaa tccccaagac ctcaaaggat ccttgcagga
     2221 gttgaaggag agggctctga gccgatacaa cctcgtgcgg ggccagggtc cagagaggct
     2281 ggtgtctggc tccgacgact tcaccttatt cctgtggtcc ccagcagagg acaaaaagcc
     2341 tctcactcgg atgacaggac accaagctct catcaaccag gtgctcttct ctcctgactc
     2401 ccgcatcgtg gctagtgcct cctttgacaa gtccatcaag ctgtgggatg gcaggacggg
     2461 caagtacctg gcttccctac gcggccacgt ggctgccgtg taccagattg cgtggtcagc
     2521 tgacagtcgg ctcctggtca gcggcagcag tgacagcaca ctgaaggtgt gggatgtgaa
     2581 ggcccagaag ctggccatgg acctgcccgg ccacgcggat gaggtatatg ctgttgactg
     2641 gagtccagat ggccagagag tggcaagtgg tgggaaggac aaatgcctcc ggatatggag
     2701 gagataatct agaggtcaag cttcgggggt ttaaacctgc aggcatgcaa gctgatttgg
     2761 ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2821 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2881 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2941 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3001 ctacaaatgt ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta
     3061 tacgaagtta tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     3121 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     3181 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3241 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3301 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3361 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3421 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3481 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3541 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3601 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3661 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3721 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3781 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3841 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3901 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa
     3961 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     4021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4681 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4801 agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     4861 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     4921 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     4981 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     5041 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     5101 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     5161 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     5221 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     5281 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     5341 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     5401 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     5461 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     5521 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     5581 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5641 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     5701 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
