LOCUS       sequence-426019-        5355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DNAAF10. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5355)
  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 5355)
  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_DNAAF10:1-357
FEATURES             Location/Qualifiers
     source          1..5355
                     /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    2418..2539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2429..2448)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2654..2687)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2697..2744
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2776..3570
                     /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(2830..2849)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3440..3459
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3728..4316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3808..3827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4442..4490
                     /note="synthetic polyadenylation signal"
     CDS             complement(4514..4912)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4909..4918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4943..5300)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4957..5092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5109..5300
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5257..5277
                     /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 tgtcggcctt
     1261 cgagaagcct cagatcatcg cccatatcca gaagggcttc aactacacgg tgtttgactg
     1321 taagtgggtg ccctgcagcg ccaaatttgt gaccatgggc aacttcgcac ggggcaccgg
     1381 cgtcattcag ctgtacgaga tccagcacgg ggacctgaag ctgcttcggg agattgaaaa
     1441 ggccaaacct attaaatgtg gaacatttgg tgcaacatct ttacagcaga gatatttagc
     1501 tactggagat tttggtggaa accttcatat atggaattta gaagctccag agatgccagt
     1561 atattctgta aagggccata aagaaattat aaatgccata gatggcatag gtggactagg
     1621 aattggagaa ggagcacctg aaattgtgac tggcagccga gatggaactg tgaaggtgtg
     1681 ggacccaagg caaaaagatg atcctgttgc taatatggaa cctgtacaag gagaaaacaa
     1741 gagagactgt tggactgtgg catttggcaa tgcttataat caagaagaac gtgttgtttg
     1801 tgctggctat gacaatgggg atatcaaact atttgatctc agaaatatgg cattacggtg
     1861 ggagacaaac atcaaaaatg gggtgtgtag cttggagttt gacagaaaag acataagtat
     1921 gaataagtta gtagccacat ctctggaagg aaagttccat gtttttgaca tgagaacaca
     1981 gcatccaacc aaaggttttg cctctgtttc agaaaaggct cataaatcta ctgtgtggca
     2041 ggtccgacac ctgccgcaga acagggagct ctttctgaca gctggaggcg ccggcggcct
     2101 tcacctctgg aagtatgaat accctattca gcggtcaaag aaagattctg agggaataga
     2161 aatgggagtc gcaggttctg taagccttct gcagaatgtt acgttgtcca cccagcccat
     2221 ttcaagtttg gattggagtc cagataaaag gggtctctgc gtctgtagtt catttgacca
     2281 aacggtgaga gtactgatcg ttacaaagct caataaaatt taatctagag gtcaagcttc
     2341 gggggtttaa acctgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc
     2401 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2461 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2521 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2581 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat
     2641 taagacttct aatataactt cgtatagcat acattatacg aagttatggt tccactgttg
     2701 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     2761 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2821 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2881 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     2941 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3001 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3061 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3121 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3181 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3241 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3301 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3361 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3421 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3481 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3541 ttctatcgcc ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac
     3601 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3661 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3721 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3781 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3841 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3901 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3961 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4021 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4081 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4141 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4201 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4261 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     4321 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4381 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg
     4441 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg
     4501 tgatggctag agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa
     4561 ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga
     4621 gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg
     4681 cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt
     4741 ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc
     4801 tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag
     4861 ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc
     4921 tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     4981 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     5041 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     5101 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     5161 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
     5221 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     5281 gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag
     5341 gccatggtgc tgcgc
//
