LOCUS       sequence-423440-        7079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human NSMAF. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7079)
  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 7079)
  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_NSMAF:1-917
FEATURES             Location/Qualifiers
     source          1..7079
                     /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    4142..4263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4153..4172)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4378..4411)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4421..4468
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4500..5294
                     /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(4554..4573)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5164..5183
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5452..6040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5532..5551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6166..6214
                     /note="synthetic polyadenylation signal"
     CDS             complement(6238..6636)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6633..6642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6667..7024)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6681..6816
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6833..7024
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6981..7001
                     /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 ccatggcgtt tatccggaag
     1261 aagcagcagg agcagcagct gcagctctac tccaaggaga gattttcctt gctgctgctt
     1321 aacttggagg agtactactt tgaacagcat agagccaatc acattttgca caagggcagt
     1381 caccatgaaa ggaaaatcag aggctcctta aaaatatgtt caaaatcggt gatttttgaa
     1441 ccagattcaa tatcccagcc catcatcaag attcctttga gagactgtat aaaaatagga
     1501 aagcatggag aaaatggagc caatagacac ttcacaaagg caaaatctgg gggtatttca
     1561 ctcattttca gtcaggtata tttcattaaa gaacataatg ttgttgcacc atataaaata
     1621 gaaaggggca aaatggaata tgtttttgaa ttggatgttc ccgggaaagt ggaagatgtt
     1681 gtggagacgt tgcttcagct tcacagagca tcctgccttg acaaattggg tgaccaaacc
     1741 gccatgataa cagctatttt gcagtctcgt ttagctagaa catcatttga caaaaacagg
     1801 ttccaaaaca tttctgaaaa gctgcacatg gaatgcaaag cagaaatggt gacgcctctg
     1861 gtgactaatc ctggacacgt gtgcatcacg gacacaaacc tgtattttca gcccctcaac
     1921 ggctacccga aacctgtggt ccagataaca ctccaagatg tccgccgcat ctacaaaagg
     1981 aggcacggcc tcatgcctct gggcttggaa gtattttgca cagaagatga tctgtgttcc
     2041 gacatctacc taaagttcta tgaacctcaa gatagagatg atctctattt ttacattgcc
     2101 acatacctag agcaccatgt ggcggagcac actgctgaga gctacatgct gcagtggcag
     2161 cgtggacacc tttccaacta tcagtacctc cttcacctca acaacctggc cgaccgcagc
     2221 tgcaacgacc tctcccagta ccctgtgttt ccatggataa tacatgatta ttccagctca
     2281 gaactagatt tgtcaaatcc aggaaccttc cgggatctca gtaagccagt aggggcccta
     2341 aataaggaac ggctggagag actactgaca cgctaccagg aaatgcctga accaaagttc
     2401 atgtatggga gtcactactc ttccccgggt tatgtacttt tttatcttgt taggattgca
     2461 ccagagtata tgctgtgcct gcagaatgga agatttgata atgcagatag aatgttcaac
     2521 agtattgcag aaacttggaa aaactgtctg gatggtgcaa cggattttaa agagttaatt
     2581 ccagaattct atggtgatga tgtgagcttt ctagtcaata gcctgaagtt ggatttggga
     2641 aagagacaag gaggacagat ggttgacgat gtggagcttc ccccttgggc ttccagtccc
     2701 gaggactttc tccagaagag caaagatgca ttggaaagca attatgtgtc tgaacacctt
     2761 cacgagtgga ttgatctaat atttggctac aaacaaaaag ggagtgatgc agttggggcc
     2821 cataatgtat ttcatcccct gacctatgaa ggaggtgtag acttgaacag catccaggat
     2881 cctgatgaga aggtagccat gcttacgcaa atcttggaat ttgggcagac accaaaacaa
     2941 ctatttgtga caccacatcc tcgaaggatc accccaaagt ttaaaagttt gtcccagacc
     3001 tccagttata atgcttctat ggcagattcc ccaggtgaag agtcttttga agacctgacc
     3061 gaagaaagca aaacactggc ctggaataac atcaccaaac tgcagttaca cgagcactat
     3121 aaaatccaca aagaagcagt tactggaatc acggtctctc gcaatggatc ttcagtattc
     3181 acaacatccc aagattccac cttgaagatg ttttctaaag aatcaaaaat gctacaaaga
     3241 agtatatcat tttcaaatat ggctttatcg tcttgtttac ttttaccagg agatgccact
     3301 gtcataactt cttcatggga taataatgtc tatttttatt ccatagcatt tggaagacgc
     3361 caggacacgt taatgggaca tgatgatgct gttagtaaga tctgttggca tgacaacagg
     3421 ctatattctg catcgtggga ctctacagtg aaggtgtggt ctggtgttcc tgcagagatg
     3481 ccaggcacca aaagacacca ctttgacttg ctggccgagc tggaacatga tgtcagtgta
     3541 gatacaatca gtttaaatgc tgcaagcaca ctgttagttt ccggcaccaa agaaggcaca
     3601 gtgaatattt gggacctcac aacggccacc ttaatgcacc agattccatg ccattcaggg
     3661 attgtatgtg acactgcttt tagcccagat agtcgccatg tcctcagcac aggaacagat
     3721 ggctgtctta atgtcattga tgtgcagaca ggaatgctca tctcctccat gacatcagat
     3781 gagccccaga ggtgctttgt ctgggatgga aattccgttt tatctggcag tcagtctggt
     3841 gaactgctcg tttgggacct ccttggagca aaaatcagtg agagaataca gggccacaca
     3901 ggtgctgtga catgtatatg gatgaatgaa cagtgtagca gtatcatcac aggaggggaa
     3961 gacagacaaa ttatattctg gaaattgcag tatggctcga gcggtgggag ctccggtgtg
     4021 agcggctggc ggctgttcaa gaagattagc taagtttaaa cggccgcgac tctagactgc
     4081 aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg agcagacatg
     4141 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4201 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4261 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     4321 tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac ttctaatata
     4381 acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt aatcatcggc
     4441 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     4501 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4561 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4621 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4681 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4741 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4801 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4861 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4921 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4981 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5041 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5101 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5161 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5221 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5281 acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     5341 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5401 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5461 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5521 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5581 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5641 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5701 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5761 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5821 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5881 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5941 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6001 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     6061 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6121 atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac aaaaaaccaa
     6181 cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagttta
     6241 gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc
     6301 atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca
     6361 ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag
     6421 gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc
     6481 agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct
     6541 ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat
     6601 gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt cagaagaatc
     6661 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     6721 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     6781 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     6841 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     6901 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     6961 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     7021 tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
