LOCUS       sequence-425864-        6545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human GEMIN5 fragment M1 to A739. C-terminal
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6545)
  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 6545)
  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_GEMIN5:1-739
FEATURES             Location/Qualifiers
     source          1..6545
                     /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    3608..3729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3619..3638)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3844..3877)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3887..3934
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3966..4760
                     /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(4020..4039)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4630..4649
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4918..5506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4998..5017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5632..5680
                     /note="synthetic polyadenylation signal"
     CDS             complement(5704..6102)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6099..6108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6133..6490)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6147..6282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6299..6490
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6447..6467
                     /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 ccatggggca ggagccgcgg
     1261 acgctgccgc cctcccccaa ctggtactgc gcccgctgca gcgatgccgt gcccgggggc
     1321 ctctttggct tcgccgcgcg gacctccgtc ttccttgtcc gcgtgggccc gggcgcaggc
     1381 gagagtccag ggacaccccc gtttcgagtc ataggagagt tggtgggaca caccgaaagg
     1441 gtctctggct tcacattttc tcatcaccct ggtcagtaca acctctgtgc caccagctcc
     1501 gacgatggga ctgtgaaaat atgggatgta gagacaaaaa cagttgtgac agaacatgca
     1561 ctccatcagc atacgatatc aacattacat tggtctcctc gagtaaagga cttaatagta
     1621 tctggggatg aaaaaggagt agttttctgt tactggttta acagaaatga cagccagcac
     1681 ctctttatag aacccaggac aattttctgt cttacttgtt cacctcatca tgaagattta
     1741 gtagccattg gctacaagga tggcatagtg gtgataattg acatcagtaa gaaaggagaa
     1801 gttattcata ggcttcgagg ccatgatgat gaaatccact ccatagcctg gtgtcccctg
     1861 cctggtgaag attgtttatc tataaaccaa gaggaaactt cagaagaagc tgaaattacc
     1921 aacgggaatg ctgtagcaca agctccagta acaaaaggtt gctacttagc cactggaagc
     1981 aaagatcaaa ccattcgaat ctggagctgt tctagaggcc gaggggtgat gattttgaaa
     2041 ttgccctttc tgaagagaag aggagggggt atagacccaa ctgttaaaga gcgcctttgg
     2101 ttgacactcc attggcccag caatcaacca acacagctgg tatctagctg ttttggaggt
     2161 gaactgttgc aatgggatct cactcaatct tggagacgga aatacaccct cttcagtgcc
     2221 tcatcagaag ggcaaaatca ttcaagaatt gtgtttaatt tatgtccttt acaaacagag
     2281 gatgacaaac agctgttact ttctacatca atggatagag atgtaaaatg ttgggacata
     2341 gccaccttgg agtgcagctg gacccttcct tcccttggtg ggtttgcata cagcctggct
     2401 ttctcttctg tggacatagg ctctttggcc ataggtgttg gggatggcat gatccgtgta
     2461 tggaatacac tctccataaa gaacaactat gatgtgaaaa atttttggca aggcgtgaag
     2521 tccaaggtta cagcgctgtg ctggcaccca accaaggaag gttgcttagc ttttggaact
     2581 gatgatggaa aagtgggatt gtatgacacc tactccaaca agcctccaca gatttctagc
     2641 acatatcata agaagactgt atatacgtta gcctgggggc caccagtacc ccccatgtca
     2701 cttggaggag aaggagacag accttccctt gctttataca gctgtggagg agaagggatt
     2761 gtcttacagc ataatccctg gaagcttagt ggagaagcct ttgacatcaa caaactcatc
     2821 agggacacca attcaatcaa atacaaattg cctgtacaca cagagataag ttggaaagca
     2881 gatggcaaaa tcatggctct tggcaatgaa gatggatcaa tagaaatatt tcagattccc
     2941 aacctgaaac tgatctgtac tatccaacag catcacaagc ttgtgaatac cattagctgg
     3001 catcatgagc atggcagcca gccagaattg agctatctga tggcctctgg ctccaacaat
     3061 gcagtcattt acgtgcacaa cctgaagact gtcatagaga gcagccctga gtctccagtg
     3121 accattacag agccctaccg gaccctctca gggcatacgg ccaagattac cagtgtggcg
     3181 tggagcccac atcatgatgg aaggctggta tctgcttcct atgatggtac agcccaggtg
     3241 tgggatgctc tccgggaaga gcccctgtgc aatttccgag gacatcaagg tcgactgctt
     3301 tgtgtggcat ggtctccttt ggatccagac tgcatctatt caggggcaga tgacttttgt
     3361 gtgcacaagt ggctcacttc catgcaagat cattcccggc ctcctcaagg caaaaaaagt
     3421 attgaattgg agaaaaaacg gctctctcaa cctaaggcag gctcgagcgg tgggagctcc
     3481 ggtgtgagcg gctggcggct gttcaagaag attagctaag tttaaacggc cgcgactcta
     3541 gactgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc gcttcgagca
     3601 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3661 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3721 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3781 gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat taagacttct
     3841 aatataactt cgtatagcat acattatacg aagttatggt tccactgttg acaattaatc
     3901 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc
     3961 agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4021 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4081 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4141 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4201 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4261 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4321 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4381 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4441 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4501 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4561 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4621 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4681 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4741 ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc
     4801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5341 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     5521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5581 gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg tcacacaaaa
     5641 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggctag
     5701 agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg
     5761 ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga gaagcacctg
     5821 tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg cgacgataca
     5881 agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt ctgcacaagg
     5941 tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc tagagagagc
     6001 tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag ccgttgctct
     6061 ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc tagtgtcaga
     6121 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     6181 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     6241 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     6301 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     6361 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     6421 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     6481 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     6541 tgcgc
//
