LOCUS       sequence-426007-        6501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human GEMIN5 fragment M1 to A739. N-terminal
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6501)
  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 6501)
  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_GEMIN5:1-739
FEATURES             Location/Qualifiers
     source          1..6501
                     /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    3564..3685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3575..3594)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3800..3833)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3843..3890
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3922..4716
                     /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(3976..3995)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4586..4605
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4874..5462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4954..4973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5588..5636
                     /note="synthetic polyadenylation signal"
     CDS             complement(5660..6058)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6055..6064
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6089..6446)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6103..6238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6255..6446
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6403..6423
                     /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 tggggcagga
     1261 gccgcggacg ctgccgccct cccccaactg gtactgcgcc cgctgcagcg atgccgtgcc
     1321 cgggggcctc tttggcttcg ccgcgcggac ctccgtcttc cttgtccgcg tgggcccggg
     1381 cgcaggcgag agtccaggga cacccccgtt tcgagtcata ggagagttgg tgggacacac
     1441 cgaaagggtc tctggcttca cattttctca tcaccctggt cagtacaacc tctgtgccac
     1501 cagctccgac gatgggactg tgaaaatatg ggatgtagag acaaaaacag ttgtgacaga
     1561 acatgcactc catcagcata cgatatcaac attacattgg tctcctcgag taaaggactt
     1621 aatagtatct ggggatgaaa aaggagtagt tttctgttac tggtttaaca gaaatgacag
     1681 ccagcacctc tttatagaac ccaggacaat tttctgtctt acttgttcac ctcatcatga
     1741 agatttagta gccattggct acaaggatgg catagtggtg ataattgaca tcagtaagaa
     1801 aggagaagtt attcataggc ttcgaggcca tgatgatgaa atccactcca tagcctggtg
     1861 tcccctgcct ggtgaagatt gtttatctat aaaccaagag gaaacttcag aagaagctga
     1921 aattaccaac gggaatgctg tagcacaagc tccagtaaca aaaggttgct acttagccac
     1981 tggaagcaaa gatcaaacca ttcgaatctg gagctgttct agaggccgag gggtgatgat
     2041 tttgaaattg ccctttctga agagaagagg agggggtata gacccaactg ttaaagagcg
     2101 cctttggttg acactccatt ggcccagcaa tcaaccaaca cagctggtat ctagctgttt
     2161 tggaggtgaa ctgttgcaat gggatctcac tcaatcttgg agacggaaat acaccctctt
     2221 cagtgcctca tcagaagggc aaaatcattc aagaattgtg tttaatttat gtcctttaca
     2281 aacagaggat gacaaacagc tgttactttc tacatcaatg gatagagatg taaaatgttg
     2341 ggacatagcc accttggagt gcagctggac ccttccttcc cttggtgggt ttgcatacag
     2401 cctggctttc tcttctgtgg acataggctc tttggccata ggtgttgggg atggcatgat
     2461 ccgtgtatgg aatacactct ccataaagaa caactatgat gtgaaaaatt tttggcaagg
     2521 cgtgaagtcc aaggttacag cgctgtgctg gcacccaacc aaggaaggtt gcttagcttt
     2581 tggaactgat gatggaaaag tgggattgta tgacacctac tccaacaagc ctccacagat
     2641 ttctagcaca tatcataaga agactgtata tacgttagcc tgggggccac cagtaccccc
     2701 catgtcactt ggaggagaag gagacagacc ttcccttgct ttatacagct gtggaggaga
     2761 agggattgtc ttacagcata atccctggaa gcttagtgga gaagcctttg acatcaacaa
     2821 actcatcagg gacaccaatt caatcaaata caaattgcct gtacacacag agataagttg
     2881 gaaagcagat ggcaaaatca tggctcttgg caatgaagat ggatcaatag aaatatttca
     2941 gattcccaac ctgaaactga tctgtactat ccaacagcat cacaagcttg tgaataccat
     3001 tagctggcat catgagcatg gcagccagcc agaattgagc tatctgatgg cctctggctc
     3061 caacaatgca gtcatttacg tgcacaacct gaagactgtc atagagagca gccctgagtc
     3121 tccagtgacc attacagagc cctaccggac cctctcaggg catacggcca agattaccag
     3181 tgtggcgtgg agcccacatc atgatggaag gctggtatct gcttcctatg atggtacagc
     3241 ccaggtgtgg gatgctctcc gggaagagcc cctgtgcaat ttccgaggac atcaaggtcg
     3301 actgctttgt gtggcatggt ctcctttgga tccagactgc atctattcag gggcagatga
     3361 cttttgtgtg cacaagtggc tcacttccat gcaagatcat tcccggcctc ctcaaggcaa
     3421 aaaaagtatt gaattggaga aaaaacggct ctctcaacct aaggcataat ctagaggtca
     3481 agcttcgggg gtttaaacct gcaggcatgc aagctgattt ggctgctgcc accgctgagc
     3541 aaggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3601 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3661 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3721 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3781 cttaattaag acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca
     3841 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3901 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3961 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4021 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4081 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4141 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4201 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4261 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4321 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4381 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4441 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4501 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4561 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4621 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4681 atcgccttct atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg
     4741 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4801 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4861 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4921 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4981 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5041 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5101 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5161 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5221 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5281 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5341 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5401 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     5461 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5521 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     5581 gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     5641 ccatcgtgat ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa
     5701 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
     5761 gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc
     5821 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
     5881 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
     5941 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
     6001 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg
     6061 tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc
     6121 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     6181 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     6241 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     6301 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     6361 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     6421 tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta
     6481 tttcaggcca tggtgctgcg c
//
