LOCUS       sequence-423439-        6392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human NOL10. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6392)
  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 6392)
  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_NOL10:1-688
FEATURES             Location/Qualifiers
     source          1..6392
                     /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             1828..1839
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3455..3576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3466..3485)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3691..3724)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3734..3781
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3813..4607
                     /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(3867..3886)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4477..4496
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4765..5353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4845..4864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5479..5527
                     /note="synthetic polyadenylation signal"
     CDS             complement(5551..5949)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5946..5955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5980..6337)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5994..6129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6146..6337
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6294..6314
                     /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 ccatgcaggt ctccagcctc
     1261 aatgaggtga agatttacag cctcagctgc ggcaagtccc ttcctgagtg gctttctgat
     1321 aggaagaaga gagcgctaca gaagaaagat gtagatgtcc gtaggagaat tgaacttatt
     1381 caggactttg aaatgcctac tgtgtgtacc actattaagg tgtcaaaaga tggacagtac
     1441 attttagcaa ctggaacata taaacctcgg gttcgatgtt atgacaccta tcaattatcc
     1501 ttgaagtttg aaaggtgttt agattcagaa gttgtcacct ttgaaatttt gtctgatgac
     1561 tactcaaaga ttgtcttctt acataatgat agatacattg aatttcattc gcaatcaggt
     1621 ttttactaca aaaccagaat accaaagttt gggagagatt tctcttacca ctatccatcc
     1681 tgtgacttgt actttgttgg tgcaagttct gaagtttata ggttaaactt agaacaagga
     1741 cgatacctga atcctctaca aactgatgct gcggagaata atgtttgtga cataaattca
     1801 gtgcatggct tgtttgccac aggaaccata gagggtagag tggagtgctg ggacccaaga
     1861 actcgaaaca gagttggcct gttagactgc gccttaaaca gtgtcacagc agattcagag
     1921 ataaacagtt taccaacaat ctctgctttg aaatttaatg gtgccttgac catggcagtt
     1981 ggaacaacca cagggcaggt tttattatat gaccttcgat ctgataagcc attgctagtt
     2041 aaagatcacc agtatgggct gcccattaag tccgttcatt tccaggattc attagatctg
     2101 attttgtctg ctgactctcg aattgtcaag atgtggaata agaactccgg aaaaatattt
     2161 acttccttgg agccagagca tgaccttaat gatgtttgtc tctaccccaa ctcaggcatg
     2221 cttctgacgg ccaatgaaac ccccaagatg ggcatctatt acattccagt tttgggtcct
     2281 gctcctcggt ggtgttcctt cttagacaac ttgaccgaag aattagaaga gaatccagaa
     2341 agcacagtct atgatgatta taaatttgtc accaagaaag accttgaaaa tttagggctc
     2401 acccacctca ttggatctcc tttcctccgg gcatatatgc atgggttttt catggatata
     2461 agactctatc acaaggtgaa actgatggta aatccatttg cttatgaaga atataggaaa
     2521 gataaaatac gacagaaaat agaagaaaca cgtgcacaga gagtccagtt aaagaaattg
     2581 ccaaaagtta acaaagagct ggcacttaaa ttaattgagg aagaagagga gaagcagaaa
     2641 tctacatgga aaaagaaagt taagagtctt cctaatattc tcaccgatga tcgatttaaa
     2701 gttatgtttg agaaccctga cttccaagta gatgaagaga gtgaagaatt taggcttctg
     2761 aatccacttg tttcaaaaat tagtgaaaaa aggaagaaga aactaagact cttagagcaa
     2821 caagaacttc gtgaaaaaga agaggaggaa gagccggaag gaaaaccaag tgatgcagaa
     2881 agttcggaga gttcagatga tgaaaaagcc tgggttgaag aggtcaggaa gcaacgcaga
     2941 ctcctccagc aggaggaaaa agtgaagcgg caggaacgac tcaaggagga ccagcagaca
     3001 gtcctaaagc cccagtttta tgagatcaaa gcaggagaag aatttagaag cttcaaagat
     3061 tctgccacaa agcagaaact gatgaacaaa acccttgaag atcgtttgaa aattgaagca
     3121 aaaaatggga cattgagtgt atccgacacc accgttggca gcaaacaatt gacattcacg
     3181 ttaaagaggt ctgaacagca gaagaagcaa caggaggctg agaaactgca tcgacaagaa
     3241 aggaaaagac tccgtcgttc ggccggacac ctgaagtcaa gacacaaaag aggacggtcg
     3301 tttcatggct cgagcggtgg gagctccggt gtgagcggct ggcggctgtt caagaagatt
     3361 agctaagttt aaacggccgc gactctagac tgcaggcatg caagctgatt tggctgctgc
     3421 caccgctgag caaggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     3481 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     3541 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3601 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     3661 tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca ttatacgaag
     3721 ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3781 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     3841 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3901 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3961 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4021 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4081 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4141 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4201 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4261 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4321 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4381 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4441 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4501 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4561 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaatgaccg
     4621 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4681 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5341 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5461 tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa aataaagata
     5521 ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac cagagggcag
     5581 caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta tggctttgat
     5641 cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca agatgcccct
     5701 gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag cagcagcagt
     5761 gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat tgacaccagt
     5821 gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt cagagatggg
     5881 gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt gagacaaagg
     5941 cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     6001 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     6061 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     6121 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     6181 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     6241 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     6301 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct
     6361 ttcagaggtt atttcaggcc atggtgctgc gc
//
