LOCUS       sequence-425934-        7227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human EML4. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7227)
  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 7227)
  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_EML4:1-981
FEATURES             Location/Qualifiers
     source          1..7227
                     /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    4290..4411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4301..4320)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4526..4559)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4569..4616
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4648..5442
                     /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(4702..4721)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5312..5331
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5600..6188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5680..5699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6314..6362
                     /note="synthetic polyadenylation signal"
     CDS             complement(6386..6784)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6781..6790
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6815..7172)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6829..6964
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6981..7172
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7129..7149
                     /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 tggacggttt
     1261 cgccggcagt ctcgatgata gtatttctgc tgcaagtact tctgatgttc aagatcgcct
     1321 gtcagctctt gagtcacgag ttcagcaaca agaagatgaa atcactgtgc taaaggcggc
     1381 tttggctgat gttttgaggc gtcttgcaat ctctgaagat catgtggcct cagtgaaaaa
     1441 atcagtctca agtaaaggcc aaccaagccc tcgagcagtt attcccatgt cctgtataac
     1501 caatggaagt ggtgcaaaca gaaaaccaag tcataccagt gctgtctcaa ttgcaggaaa
     1561 agaaactctt tcatctgctg ctaaaagtgg tacagaaaaa aagaaagaaa aaccacaagg
     1621 acagagagaa aaaaaagagg aatctcattc taatgatcaa agtccacaaa ttcgagcatc
     1681 accttctccc cagccctctt cacaacctct ccaaatacac agacaaactc cagaaagcaa
     1741 gaatgctact cccaccaaaa gcataaaacg accatcacca gctgaaaagt cacataattc
     1801 ttgggaaaat tcagatgata gccgtaataa attgtcgaaa ataccttcaa cacccaaatt
     1861 aataccaaaa gttaccaaaa ctgcagacaa gcataaagat gtcatcatca accaagaagg
     1921 agaatatatt aaaatgttta tgcgcggtcg gccaattacc atgttcattc cttccgatgt
     1981 tgacaactat gatgacatca gaacggaact gcctcctgag aagctcaaac tggagtgggc
     2041 atatggttat cgaggaaagg actgtagagc taatgtttac cttcttccga ccggggaaat
     2101 agtttatttc attgcatcag tagtagtact atttaattat gaggagagaa ctcagcgaca
     2161 ctacctgggc catacagact gtgtgaaatg ccttgctata catcctgaca aaattaggat
     2221 tgcaactgga cagatagctg gcgtggataa agatggaagg cctctacaac cccacgtcag
     2281 agtgtgggat tctgttactc tatccacact gcagattatt ggacttggca cttttgagcg
     2341 tggagtagga tgcctggatt tttcaaaagc agattcaggt gttcatttat gtgttattga
     2401 tgactccaat gagcatatgc ttactgtatg ggactggcag aagaaagcaa aaggagcaga
     2461 aataaagaca acaaatgaag ttgttttggc tgtggagttt cacccaacag atgcaaatac
     2521 cataattaca tgcggtaaat ctcatatttt cttctggacc tggagcggca attcactaac
     2581 aagaaaacag ggaatttttg ggaaatatga aaagccaaaa tttgtgcagt gtttagcatt
     2641 cttggggaat ggagatgttc ttactggaga ctcaggtgga gtcatgctta tatggagcaa
     2701 aactactgta gagcccacac ctgggaaagg acctaaaggt gtatatcaaa tcagcaaaca
     2761 aatcaaagct catgatggca gtgtgttcac actttgtcag atgagaaatg ggatgttatt
     2821 aactggagga gggaaagaca gaaaaataat tctgtgggat catgatctga atcctgaaag
     2881 agaaatagag gttcctgatc agtatggcac aatcagagct gtagcagaag gaaaggcaga
     2941 tcaattttta gtaggcacat cacgaaactt tattttacga ggaacattta atgatggctt
     3001 ccaaatagaa gtacagggtc atacagatga gctttggggt cttgccacac atcccttcaa
     3061 agatttgctc ttgacatgtg ctcaggacag gcaggtgtgc ctgtggaact caatggaaca
     3121 caggctggaa tggaccaggc tggtagatga accaggacac tgtgcagatt ttcatccaag
     3181 tggcacagtg gtggccatag gaacgcactc aggcaggtgg tttgttctgg atgcagaaac
     3241 cagagatcta gtttctatcc acacagacgg gaatgaacag ctctctgtga tgcgctactc
     3301 aatagatggt accttcctgg ctgtaggatc tcatgacaac tttatttacc tctatgtagt
     3361 ctctgaaaat ggaagaaaat atagcagata tggaaggtgc actggacatt ccagctacat
     3421 cacacacctt gactggtccc cagacaacaa gtatataatg tctaactcgg gagactatga
     3481 aatattgtac tgggacattc caaatggctg caaactaatc aggaatcgat cggattgtaa
     3541 ggacattgat tggacgacat atacctgtgt gctaggattt caagtatttg gtgtctggcc
     3601 agaaggatct gatgggacag atatcaatgc actggtgcga tcccacaata gaaaggtgat
     3661 agctgttgcc gatgactttt gtaaagtcca tctgtttcag tatccctgct ccaaagcaaa
     3721 ggctcccagt cacaagtaca gtgcccacag cagccatgtc accaatgtca gttttactca
     3781 caatgacagt cacctgatat caactggtgg aaaagacatg agcatcattc agtggaaact
     3841 tgtggaaaag ttatctttgc ctcagaatga gactgtagcg gatactactc taaccaaagc
     3901 ccccgtctct tccactgaaa gtgtcatcca atctaatact cccacaccgc ctccttctca
     3961 gcccttaaat gagacagctg aagaggaaag tagaataagc agttctccca cacttctgga
     4021 gaacagcctg gaacaaactg tggagccaag tgaagaccac agcgaggagg agagtgaaga
     4081 gggcagcgga gaccttggtg agcctcttta tgaagagcca tgcaacgaga taagcaagga
     4141 gcaggccaaa gccacccttc tggaggacca gcaagaccct tcgccctcgt cctaatctag
     4201 aggtcaagct tcgggggttt aaacctgcag gcatgcaagc tgatttggct gctgccaccg
     4261 ctgagcaagg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     4321 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4381 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4441 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     4501 taaaatctta attaagactt ctaatataac ttcgtatagc atacattata cgaagttatg
     4561 gttccactgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     4621 tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct
     4681 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4741 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4801 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4861 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4921 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4981 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5041 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5101 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5161 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5221 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5281 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5341 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5401 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gattcgaaat gaccgaccaa
     5461 gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5521 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5581 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5641 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5701 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5761 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5821 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5881 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5941 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6001 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6061 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6121 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6181 gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6241 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttat
     6301 agtccggaat cgtcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta
     6361 ttgcggccat cgtgatggct agagtttagc cctcccacac ataaccagag ggcagcaatt
     6421 cacgaatccc aactgccgtc ggctgtccat cactgtcctt cactatggct ttgatcccag
     6481 gatgcagatc gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg cccctgttct
     6541 catttccgat cgcgacgata caagtcaggt tgccagctgc cgcagcagca gcagtgccca
     6601 gcaccacgag ttctgcacaa ggtcccccag taaaatgata tacattgaca ccagtgaaga
     6661 tgcggccgtc gctagagaga gctgcgctgg cgacgctgta gtcttcagag atggggatgc
     6721 tgttgattgt agccgttgct ctttcaatga gggtggattc ttcttgagac aaaggcttgg
     6781 ccatggtggc gctagtgtca gaagaatcga gctttttgca aaagcctagg cctccaaaaa
     6841 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     6901 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     6961 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     7021 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     7081 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     7141 acacattcca cagctggttc tttccgcctc agaaggtacc taaccaagtt cctctttcag
     7201 aggttatttc aggccatggt gctgcgc
//
