LOCUS       sequence-425912-        7035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human NSMAF. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7035)
  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 7035)
  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_NSMAF:1-917
FEATURES             Location/Qualifiers
     source          1..7035
                     /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    4098..4219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4109..4128)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4334..4367)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4377..4424
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4456..5250
                     /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(4510..4529)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5120..5139
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5408..5996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5488..5507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6122..6170
                     /note="synthetic polyadenylation signal"
     CDS             complement(6194..6592)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6589..6598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6623..6980)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6637..6772
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6789..6980
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6937..6957
                     /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 tggcgtttat
     1261 ccggaagaag cagcaggagc agcagctgca gctctactcc aaggagagat tttccttgct
     1321 gctgcttaac ttggaggagt actactttga acagcataga gccaatcaca ttttgcacaa
     1381 gggcagtcac catgaaagga aaatcagagg ctccttaaaa atatgttcaa aatcggtgat
     1441 ttttgaacca gattcaatat cccagcccat catcaagatt cctttgagag actgtataaa
     1501 aataggaaag catggagaaa atggagccaa tagacacttc acaaaggcaa aatctggggg
     1561 tatttcactc attttcagtc aggtatattt cattaaagaa cataatgttg ttgcaccata
     1621 taaaatagaa aggggcaaaa tggaatatgt ttttgaattg gatgttcccg ggaaagtgga
     1681 agatgttgtg gagacgttgc ttcagcttca cagagcatcc tgccttgaca aattgggtga
     1741 ccaaaccgcc atgataacag ctattttgca gtctcgttta gctagaacat catttgacaa
     1801 aaacaggttc caaaacattt ctgaaaagct gcacatggaa tgcaaagcag aaatggtgac
     1861 gcctctggtg actaatcctg gacacgtgtg catcacggac acaaacctgt attttcagcc
     1921 cctcaacggc tacccgaaac ctgtggtcca gataacactc caagatgtcc gccgcatcta
     1981 caaaaggagg cacggcctca tgcctctggg cttggaagta ttttgcacag aagatgatct
     2041 gtgttccgac atctacctaa agttctatga acctcaagat agagatgatc tctattttta
     2101 cattgccaca tacctagagc accatgtggc ggagcacact gctgagagct acatgctgca
     2161 gtggcagcgt ggacaccttt ccaactatca gtacctcctt cacctcaaca acctggccga
     2221 ccgcagctgc aacgacctct cccagtaccc tgtgtttcca tggataatac atgattattc
     2281 cagctcagaa ctagatttgt caaatccagg aaccttccgg gatctcagta agccagtagg
     2341 ggccctaaat aaggaacggc tggagagact actgacacgc taccaggaaa tgcctgaacc
     2401 aaagttcatg tatgggagtc actactcttc cccgggttat gtactttttt atcttgttag
     2461 gattgcacca gagtatatgc tgtgcctgca gaatggaaga tttgataatg cagatagaat
     2521 gttcaacagt attgcagaaa cttggaaaaa ctgtctggat ggtgcaacgg attttaaaga
     2581 gttaattcca gaattctatg gtgatgatgt gagctttcta gtcaatagcc tgaagttgga
     2641 tttgggaaag agacaaggag gacagatggt tgacgatgtg gagcttcccc cttgggcttc
     2701 cagtcccgag gactttctcc agaagagcaa agatgcattg gaaagcaatt atgtgtctga
     2761 acaccttcac gagtggattg atctaatatt tggctacaaa caaaaaggga gtgatgcagt
     2821 tggggcccat aatgtatttc atcccctgac ctatgaagga ggtgtagact tgaacagcat
     2881 ccaggatcct gatgagaagg tagccatgct tacgcaaatc ttggaatttg ggcagacacc
     2941 aaaacaacta tttgtgacac cacatcctcg aaggatcacc ccaaagttta aaagtttgtc
     3001 ccagacctcc agttataatg cttctatggc agattcccca ggtgaagagt cttttgaaga
     3061 cctgaccgaa gaaagcaaaa cactggcctg gaataacatc accaaactgc agttacacga
     3121 gcactataaa atccacaaag aagcagttac tggaatcacg gtctctcgca atggatcttc
     3181 agtattcaca acatcccaag attccacctt gaagatgttt tctaaagaat caaaaatgct
     3241 acaaagaagt atatcatttt caaatatggc tttatcgtct tgtttacttt taccaggaga
     3301 tgccactgtc ataacttctt catgggataa taatgtctat ttttattcca tagcatttgg
     3361 aagacgccag gacacgttaa tgggacatga tgatgctgtt agtaagatct gttggcatga
     3421 caacaggcta tattctgcat cgtgggactc tacagtgaag gtgtggtctg gtgttcctgc
     3481 agagatgcca ggcaccaaaa gacaccactt tgacttgctg gccgagctgg aacatgatgt
     3541 cagtgtagat acaatcagtt taaatgctgc aagcacactg ttagtttccg gcaccaaaga
     3601 aggcacagtg aatatttggg acctcacaac ggccacctta atgcaccaga ttccatgcca
     3661 ttcagggatt gtatgtgaca ctgcttttag cccagatagt cgccatgtcc tcagcacagg
     3721 aacagatggc tgtcttaatg tcattgatgt gcagacagga atgctcatct cctccatgac
     3781 atcagatgag ccccagaggt gctttgtctg ggatggaaat tccgttttat ctggcagtca
     3841 gtctggtgaa ctgctcgttt gggacctcct tggagcaaaa atcagtgaga gaatacaggg
     3901 ccacacaggt gctgtgacat gtatatggat gaatgaacag tgtagcagta tcatcacagg
     3961 aggggaagac agacaaatta tattctggaa attgcagtat taatctagag gtcaagcttc
     4021 gggggtttaa acctgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc
     4081 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4141 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4201 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4261 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat
     4321 taagacttct aatataactt cgtatagcat acattatacg aagttatggt tccactgttg
     4381 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     4441 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4501 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4561 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4621 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4681 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4741 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4801 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4861 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4921 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4981 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5041 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5101 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5161 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5221 ttctatcgcc ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac
     5281 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5341 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5821 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     6001 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6061 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg
     6121 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg
     6181 tgatggctag agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa
     6241 ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga
     6301 gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg
     6361 cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt
     6421 ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc
     6481 tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag
     6541 ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc
     6601 tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     6661 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     6721 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     6781 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     6841 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
     6901 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     6961 gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag
     7021 gccatggtgc tgcgc
//
