LOCUS       sequence-423462-        7271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human EML4. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7271)
  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 7271)
  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_EML4:1-981
FEATURES             Location/Qualifiers
     source          1..7271
                     /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    4334..4455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4345..4364)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4570..4603)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4613..4660
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4692..5486
                     /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(4746..4765)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5356..5375
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5644..6232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5724..5743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6358..6406
                     /note="synthetic polyadenylation signal"
     CDS             complement(6430..6828)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6825..6834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6859..7216)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6873..7008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7025..7216
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7173..7193
                     /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 ccatggacgg tttcgccggc
     1261 agtctcgatg atagtatttc tgctgcaagt acttctgatg ttcaagatcg cctgtcagct
     1321 cttgagtcac gagttcagca acaagaagat gaaatcactg tgctaaaggc ggctttggct
     1381 gatgttttga ggcgtcttgc aatctctgaa gatcatgtgg cctcagtgaa aaaatcagtc
     1441 tcaagtaaag gccaaccaag ccctcgagca gttattccca tgtcctgtat aaccaatgga
     1501 agtggtgcaa acagaaaacc aagtcatacc agtgctgtct caattgcagg aaaagaaact
     1561 ctttcatctg ctgctaaaag tggtacagaa aaaaagaaag aaaaaccaca aggacagaga
     1621 gaaaaaaaag aggaatctca ttctaatgat caaagtccac aaattcgagc atcaccttct
     1681 ccccagccct cttcacaacc tctccaaata cacagacaaa ctccagaaag caagaatgct
     1741 actcccacca aaagcataaa acgaccatca ccagctgaaa agtcacataa ttcttgggaa
     1801 aattcagatg atagccgtaa taaattgtcg aaaatacctt caacacccaa attaatacca
     1861 aaagttacca aaactgcaga caagcataaa gatgtcatca tcaaccaaga aggagaatat
     1921 attaaaatgt ttatgcgcgg tcggccaatt accatgttca ttccttccga tgttgacaac
     1981 tatgatgaca tcagaacgga actgcctcct gagaagctca aactggagtg ggcatatggt
     2041 tatcgaggaa aggactgtag agctaatgtt taccttcttc cgaccgggga aatagtttat
     2101 ttcattgcat cagtagtagt actatttaat tatgaggaga gaactcagcg acactacctg
     2161 ggccatacag actgtgtgaa atgccttgct atacatcctg acaaaattag gattgcaact
     2221 ggacagatag ctggcgtgga taaagatgga aggcctctac aaccccacgt cagagtgtgg
     2281 gattctgtta ctctatccac actgcagatt attggacttg gcacttttga gcgtggagta
     2341 ggatgcctgg atttttcaaa agcagattca ggtgttcatt tatgtgttat tgatgactcc
     2401 aatgagcata tgcttactgt atgggactgg cagaagaaag caaaaggagc agaaataaag
     2461 acaacaaatg aagttgtttt ggctgtggag tttcacccaa cagatgcaaa taccataatt
     2521 acatgcggta aatctcatat tttcttctgg acctggagcg gcaattcact aacaagaaaa
     2581 cagggaattt ttgggaaata tgaaaagcca aaatttgtgc agtgtttagc attcttgggg
     2641 aatggagatg ttcttactgg agactcaggt ggagtcatgc ttatatggag caaaactact
     2701 gtagagccca cacctgggaa aggacctaaa ggtgtatatc aaatcagcaa acaaatcaaa
     2761 gctcatgatg gcagtgtgtt cacactttgt cagatgagaa atgggatgtt attaactgga
     2821 ggagggaaag acagaaaaat aattctgtgg gatcatgatc tgaatcctga aagagaaata
     2881 gaggttcctg atcagtatgg cacaatcaga gctgtagcag aaggaaaggc agatcaattt
     2941 ttagtaggca catcacgaaa ctttatttta cgaggaacat ttaatgatgg cttccaaata
     3001 gaagtacagg gtcatacaga tgagctttgg ggtcttgcca cacatccctt caaagatttg
     3061 ctcttgacat gtgctcagga caggcaggtg tgcctgtgga actcaatgga acacaggctg
     3121 gaatggacca ggctggtaga tgaaccagga cactgtgcag attttcatcc aagtggcaca
     3181 gtggtggcca taggaacgca ctcaggcagg tggtttgttc tggatgcaga aaccagagat
     3241 ctagtttcta tccacacaga cgggaatgaa cagctctctg tgatgcgcta ctcaatagat
     3301 ggtaccttcc tggctgtagg atctcatgac aactttattt acctctatgt agtctctgaa
     3361 aatggaagaa aatatagcag atatggaagg tgcactggac attccagcta catcacacac
     3421 cttgactggt ccccagacaa caagtatata atgtctaact cgggagacta tgaaatattg
     3481 tactgggaca ttccaaatgg ctgcaaacta atcaggaatc gatcggattg taaggacatt
     3541 gattggacga catatacctg tgtgctagga tttcaagtat ttggtgtctg gccagaagga
     3601 tctgatggga cagatatcaa tgcactggtg cgatcccaca atagaaaggt gatagctgtt
     3661 gccgatgact tttgtaaagt ccatctgttt cagtatccct gctccaaagc aaaggctccc
     3721 agtcacaagt acagtgccca cagcagccat gtcaccaatg tcagttttac tcacaatgac
     3781 agtcacctga tatcaactgg tggaaaagac atgagcatca ttcagtggaa acttgtggaa
     3841 aagttatctt tgcctcagaa tgagactgta gcggatacta ctctaaccaa agcccccgtc
     3901 tcttccactg aaagtgtcat ccaatctaat actcccacac cgcctccttc tcagccctta
     3961 aatgagacag ctgaagagga aagtagaata agcagttctc ccacacttct ggagaacagc
     4021 ctggaacaaa ctgtggagcc aagtgaagac cacagcgagg aggagagtga agagggcagc
     4081 ggagaccttg gtgagcctct ttatgaagag ccatgcaacg agataagcaa ggagcaggcc
     4141 aaagccaccc ttctggagga ccagcaagac ccttcgccct cgtccggctc gagcggtggg
     4201 agctccggtg tgagcggctg gcggctgttc aagaagatta gctaagttta aacggccgcg
     4261 actctagact gcaggcatgc aagctgattt ggctgctgcc accgctgagc aaggccgctt
     4321 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4381 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4441 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4501 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cttaattaag
     4561 acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca ctgttgacaa
     4621 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     4681 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4741 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4801 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4861 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4921 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4981 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5041 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5101 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5161 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5221 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5281 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5341 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5401 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5461 atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg cccaaccggt
     5521 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5581 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5641 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5701 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5761 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5821 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5881 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5941 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6001 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6061 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6121 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6181 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     6241 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6301 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaatcgtcac
     6361 acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat
     6421 ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa tcccaactgc
     6481 cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca gatcgagaag
     6541 cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc cgatcgcgac
     6601 gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca cgagttctgc
     6661 acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc cgtcgctaga
     6721 gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga ttgtagccgt
     6781 tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg tggcgctagt
     6841 gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     6901 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     6961 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     7021 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     7081 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     7141 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     7201 gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta tttcaggcca
     7261 tggtgctgcg c
//
