LOCUS       sequence-425878-        5399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DNAAF10. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5399)
  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 5399)
  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_DNAAF10:1-357
FEATURES             Location/Qualifiers
     source          1..5399
                     /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    2462..2583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2473..2492)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2698..2731)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2741..2788
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2820..3614
                     /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(2874..2893)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3484..3503
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3772..4360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3852..3871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4486..4534
                     /note="synthetic polyadenylation signal"
     CDS             complement(4558..4956)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4953..4962
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4987..5344)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5001..5136
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5153..5344
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5301..5321
                     /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 ccatgtcggc cttcgagaag
     1261 cctcagatca tcgcccatat ccagaagggc ttcaactaca cggtgtttga ctgtaagtgg
     1321 gtgccctgca gcgccaaatt tgtgaccatg ggcaacttcg cacggggcac cggcgtcatt
     1381 cagctgtacg agatccagca cggggacctg aagctgcttc gggagattga aaaggccaaa
     1441 cctattaaat gtggaacatt tggtgcaaca tctttacagc agagatattt agctactgga
     1501 gattttggtg gaaaccttca tatatggaat ttagaagctc cagagatgcc agtatattct
     1561 gtaaagggcc ataaagaaat tataaatgcc atagatggca taggtggact aggaattgga
     1621 gaaggagcac ctgaaattgt gactggcagc cgagatggaa ctgtgaaggt gtgggaccca
     1681 aggcaaaaag atgatcctgt tgctaatatg gaacctgtac aaggagaaaa caagagagac
     1741 tgttggactg tggcatttgg caatgcttat aatcaagaag aacgtgttgt ttgtgctggc
     1801 tatgacaatg gggatatcaa actatttgat ctcagaaata tggcattacg gtgggagaca
     1861 aacatcaaaa atggggtgtg tagcttggag tttgacagaa aagacataag tatgaataag
     1921 ttagtagcca catctctgga aggaaagttc catgtttttg acatgagaac acagcatcca
     1981 accaaaggtt ttgcctctgt ttcagaaaag gctcataaat ctactgtgtg gcaggtccga
     2041 cacctgccgc agaacaggga gctctttctg acagctggag gcgccggcgg ccttcacctc
     2101 tggaagtatg aataccctat tcagcggtca aagaaagatt ctgagggaat agaaatggga
     2161 gtcgcaggtt ctgtaagcct tctgcagaat gttacgttgt ccacccagcc catttcaagt
     2221 ttggattgga gtccagataa aaggggtctc tgcgtctgta gttcatttga ccaaacggtg
     2281 agagtactga tcgttacaaa gctcaataaa attggctcga gcggtgggag ctccggtgtg
     2341 agcggctggc ggctgttcaa gaagattagc taagtttaaa cggccgcgac tctagactgc
     2401 aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg agcagacatg
     2461 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2521 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2581 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2641 tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac ttctaatata
     2701 acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt aatcatcggc
     2761 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     2821 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2881 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2941 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3001 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3061 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3121 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3181 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3241 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3301 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3361 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3421 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3481 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3541 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3601 acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     3661 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3721 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3781 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3841 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3901 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3961 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4021 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4081 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4141 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4201 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4261 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4321 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     4381 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4441 atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac aaaaaaccaa
     4501 cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagttta
     4561 gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc
     4621 atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca
     4681 ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag
     4741 gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc
     4801 agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct
     4861 ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat
     4921 gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt cagaagaatc
     4981 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     5041 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     5101 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     5161 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5221 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5281 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     5341 tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
