LOCUS       sequence-423438-        5306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human MLST8. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5306)
  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 5306)
  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_MLST8:1-326
FEATURES             Location/Qualifiers
     source          1..5306
                     /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    2369..2490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2380..2399)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2605..2638)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2648..2695
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2727..3521
                     /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(2781..2800)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3391..3410
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3679..4267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3759..3778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4393..4441
                     /note="synthetic polyadenylation signal"
     CDS             complement(4465..4863)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4860..4869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4894..5251)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4908..5043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5060..5251
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5208..5228
                     /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 ccatgaacac ctccccaggc
     1261 acggtgggca gtgacccggt catcctggcc actgcaggct acgaccacac cgtgcgcttc
     1321 tggcaggccc acagcggcat ctgcacccgg acggtgcagc accaggactc ccaggtgaat
     1381 gccttggagg tcacaccgga ccgcagcatg attgctgctg caggttacca gcacatccgc
     1441 atgtatgatc tcaactccaa taaccctaac cccatcatca gctacgacgg cgtcaacaag
     1501 aacatcgcgt ctgtgggctt ccacgaagac ggccgctgga tgtacacggg cggcgaggac
     1561 tgcacagcca ggatctggga cctcaggtcc cggaacctgc agtgccagcg gatcttccag
     1621 gtgaacgcac ccattaactg cgtgtgcctg caccccaacc aggcagagct catcgtgggt
     1681 gaccagagcg gggctatcca catctgggac ttgaaaacag accacaacga gcagctgatc
     1741 cctgagcccg aggtctccat cacgtccgcc cacatcgatc ccgacgccag ctacatggca
     1801 gctgtcaata gcaccggaaa ctgctatgtc tggaatctga cggggggcat tggtgacgag
     1861 gtgacccagc tcatccccaa gactaagatc cctgcccaca cgcgctacgc cctgcagtgt
     1921 cgcttcagcc ccgactccac gctcctcgcc acctgctcgg ctgatcagac gtgcaagatc
     1981 tggaggacgt ccaacttctc cctgatgacg gagctgagca tcaagagcgg caaccccggg
     2041 gagtcctccc gcggctggat gtggggctgc gccttctcgg gggactccca gtacatcgtc
     2101 actgcttcct cggacaacct ggcccggctc tggtgtgtgg agactggaga gatcaagaga
     2161 gagtatggcg gccaccagaa ggctgttgtc tgcctggcct tcaatgacag tgtgctgggc
     2221 ggctcgagcg gtgggagctc cggtgtgagc ggctggcggc tgttcaagaa gattagctaa
     2281 gtttaaacgg ccgcgactct agactgcagg catgcaagct gatttggctg ctgccaccgc
     2341 tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2401 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2461 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2521 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     2581 aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac gaagttatgg
     2641 ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     2701 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     2761 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2821 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2881 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     2941 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3001 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3061 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3121 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3181 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3241 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3301 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3361 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3421 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3481 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     3541 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3601 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3661 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3721 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3781 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3841 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3901 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3961 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4021 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4081 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4141 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4201 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4261 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4321 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     4381 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     4441 tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg gcagcaattc
     4501 acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg
     4561 atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc
     4621 atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag
     4681 caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat
     4741 gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct
     4801 gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc
     4861 catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa
     4921 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     4981 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     5041 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     5101 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     5161 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     5221 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
     5281 ggttatttca ggccatggtg ctgcgc
//
