LOCUS       sequence-425911-        6348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human NOL10. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6348)
  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 6348)
  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_NOL10:1-688
FEATURES             Location/Qualifiers
     source          1..6348
                     /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"
     CDS             1835..1846
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3411..3532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3422..3441)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3647..3680)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3690..3737
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3769..4563
                     /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(3823..3842)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4433..4452
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4721..5309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4801..4820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5435..5483
                     /note="synthetic polyadenylation signal"
     CDS             complement(5507..5905)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5902..5911
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5936..6293)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5950..6085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6102..6293
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6250..6270
                     /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 tgcaggtctc
     1261 cagcctcaat gaggtgaaga tttacagcct cagctgcggc aagtcccttc ctgagtggct
     1321 ttctgatagg aagaagagag cgctacagaa gaaagatgta gatgtccgta ggagaattga
     1381 acttattcag gactttgaaa tgcctactgt gtgtaccact attaaggtgt caaaagatgg
     1441 acagtacatt ttagcaactg gaacatataa acctcgggtt cgatgttatg acacctatca
     1501 attatccttg aagtttgaaa ggtgtttaga ttcagaagtt gtcacctttg aaattttgtc
     1561 tgatgactac tcaaagattg tcttcttaca taatgataga tacattgaat ttcattcgca
     1621 atcaggtttt tactacaaaa ccagaatacc aaagtttggg agagatttct cttaccacta
     1681 tccatcctgt gacttgtact ttgttggtgc aagttctgaa gtttataggt taaacttaga
     1741 acaaggacga tacctgaatc ctctacaaac tgatgctgcg gagaataatg tttgtgacat
     1801 aaattcagtg catggcttgt ttgccacagg aaccatagag ggtagagtgg agtgctggga
     1861 cccaagaact cgaaacagag ttggcctgtt agactgcgcc ttaaacagtg tcacagcaga
     1921 ttcagagata aacagtttac caacaatctc tgctttgaaa tttaatggtg ccttgaccat
     1981 ggcagttgga acaaccacag ggcaggtttt attatatgac cttcgatctg ataagccatt
     2041 gctagttaaa gatcaccagt atgggctgcc cattaagtcc gttcatttcc aggattcatt
     2101 agatctgatt ttgtctgctg actctcgaat tgtcaagatg tggaataaga actccggaaa
     2161 aatatttact tccttggagc cagagcatga ccttaatgat gtttgtctct accccaactc
     2221 aggcatgctt ctgacggcca atgaaacccc caagatgggc atctattaca ttccagtttt
     2281 gggtcctgct cctcggtggt gttccttctt agacaacttg accgaagaat tagaagagaa
     2341 tccagaaagc acagtctatg atgattataa atttgtcacc aagaaagacc ttgaaaattt
     2401 agggctcacc cacctcattg gatctccttt cctccgggca tatatgcatg ggtttttcat
     2461 ggatataaga ctctatcaca aggtgaaact gatggtaaat ccatttgctt atgaagaata
     2521 taggaaagat aaaatacgac agaaaataga agaaacacgt gcacagagag tccagttaaa
     2581 gaaattgcca aaagttaaca aagagctggc acttaaatta attgaggaag aagaggagaa
     2641 gcagaaatct acatggaaaa agaaagttaa gagtcttcct aatattctca ccgatgatcg
     2701 atttaaagtt atgtttgaga accctgactt ccaagtagat gaagagagtg aagaatttag
     2761 gcttctgaat ccacttgttt caaaaattag tgaaaaaagg aagaagaaac taagactctt
     2821 agagcaacaa gaacttcgtg aaaaagaaga ggaggaagag ccggaaggaa aaccaagtga
     2881 tgcagaaagt tcggagagtt cagatgatga aaaagcctgg gttgaagagg tcaggaagca
     2941 acgcagactc ctccagcagg aggaaaaagt gaagcggcag gaacgactca aggaggacca
     3001 gcagacagtc ctaaagcccc agttttatga gatcaaagca ggagaagaat ttagaagctt
     3061 caaagattct gccacaaagc agaaactgat gaacaaaacc cttgaagatc gtttgaaaat
     3121 tgaagcaaaa aatgggacat tgagtgtatc cgacaccacc gttggcagca aacaattgac
     3181 attcacgtta aagaggtctg aacagcagaa gaagcaacag gaggctgaga aactgcatcg
     3241 acaagaaagg aaaagactcc gtcgttcggc cggacacctg aagtcaagac acaaaagagg
     3301 acggtcgttt cattaatcta gaggtcaagc ttcgggggtt taaacctgca ggcatgcaag
     3361 ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat
     3421 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3481 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3541 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     3601 gtaaaacctc tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag
     3661 catacattat acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg
     3721 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     3781 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3841 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3901 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3961 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4021 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4081 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4141 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4201 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4261 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4321 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4381 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4441 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4501 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4561 tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     4621 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4681 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4741 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4801 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4861 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4921 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4981 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5041 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5101 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5161 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5221 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5281 agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg
     5341 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5401 tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt
     5461 aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca
     5521 cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct
     5581 tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg
     5641 ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg
     5701 ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat
     5761 atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt
     5821 agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt
     5881 cttcttgaga caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc
     5941 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     6001 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     6061 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     6121 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     6181 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     6241 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac
     6301 ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgc
//
