LOCUS       sequence-424578-        6594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human TLE1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6594)
  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 6594)
  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_TLE1:1-770
FEATURES             Location/Qualifiers
     source          1..6594
                     /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    3657..3778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3668..3687)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3893..3926)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3936..3983
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4015..4809
                     /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(4069..4088)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4679..4698
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4967..5555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5047..5066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5681..5729
                     /note="synthetic polyadenylation signal"
     CDS             complement(5753..6151)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6148..6157
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6182..6539)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6196..6331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6348..6539
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6496..6516
                     /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 tgttcccgca
     1261 gagccggcac ccgacgccgc accaggctgc aggccagccc ttcaagttca ctatcccgga
     1321 gtccctggac cggattaaag aggaattcca gttcctgcag gcgcagtatc acagccttaa
     1381 attggaatgt gagaaactgg caagtgaaaa gacagaaatg cagaggcact atgtgatgta
     1441 ttatgaaatg tcatatggat taaacattga aatgcacaaa cagactgaaa tcgccaagag
     1501 attgaatacg atttgtgcac aagtcatccc atttctgtct caggaacatc aacaacaggt
     1561 ggcccaggct gttgaacgtg ccaaacaggt gaccatggca gagttgaatg ccatcatcgg
     1621 gcagcagcag ttgcaagctc agcatctttc tcatggccac ggacccccag ttccccttac
     1681 gcctcaccct tcgggacttc agcctcctgg aatcccgccc ctcgggggca gtgccggcct
     1741 tcttgcgctg tctagtgctc tgagtgggca gtctcacttg gcaataaaag atgacaagaa
     1801 gcaccacgat gcagagcacc acagagacag agagccgggc acaagtaatt ccctcctggt
     1861 cccagacagt ctaagaggca cagataaacg cagaaatgga cctgaatttt ccaatgacat
     1921 caagaaaagg aaggtggatg ataaggactc cagccactat gacagtgatg gtgacaaaag
     1981 cgatgacaac ttagttgtgg atgtgtctaa tgaggaccct tcttctccgc gagcaagccc
     2041 tgcccactcg ccccgggaaa atggaatcga caaaaatcgc ctgctaaaga aggatgcttc
     2101 tagcagtcca gcttccacgg cctcctcggc aagttccact tctttgaaat ccaaagaaat
     2161 gagcttgcat gaaaaagcca gcacgcctgt tctgaaatcc agcacaccaa cgcctcggag
     2221 cgacatgcca acgccgggca ccagcgccac tccaggcctc cgtccaggtc tcggcaagcc
     2281 tccagccata gaccccctcg ttaaccaagc ggcagctggc ttgaggacac ccctggcagt
     2341 gcccggccca tatcctgctc cttttgggat ggtcccccac gctggcatga acggcgagct
     2401 gaccagccca ggcgctgcct acgccagttt acacaacatg tcgccccaga tgagcgccgc
     2461 agccgccgcg gccgccgtgg tggcctacgg gcgctccccc atggtggggt ttgatcctcc
     2521 ccctcacatg agagtaccta ccattcctcc aaacctggca ggaatccctg gggggaaacc
     2581 tgcatactcc ttccacgtta ctgcagacgg tcagatgcag cctgtcccct ttccccccga
     2641 cgccctcatc ggacccggaa tcccccggca tgctcgccag atcaacaccc tcaaccacgg
     2701 ggaggtggtg tgcgctgtga ccatcagcaa ccccacgaga cacgtgtaca caggcgggaa
     2761 gggctgcgtc aaggtctggg acatcagcca ccctggcaat aagagccctg tctcccagct
     2821 cgactgtctg aacagagaca attatatccg ttcctgtaaa ttgctacccg atggctgcac
     2881 tctcatagtg ggaggggaag ccagtacttt gtccatttgg gacctggcgg ctccaacccc
     2941 gcgcatcaag gcggagctga cgtcctcggc ccccgcctgc tacgccctgg ccatcagccc
     3001 cgattccaag gtctgcttct catgctgcag cgacggcaac atcgctgtgt gggatctgca
     3061 caaccagaca ctagtgaggc aattccaggg ccacacagac ggagccagct gtattgacat
     3121 ttctaatgat ggcaccaagc tctggacggg tggtttggac aacacagtca ggtcctggga
     3181 cctgcgcgag gggcggcagc tgcagcagca cgacttcacc tcccagatct tctccctggg
     3241 gtactgcccc accggggagt ggctggcagt gggcatggag agcagcaatg tggaggtgct
     3301 gcacgtgaac aagcctgaca agtaccagct gcacctgcat gagagctgcg tgctgtccct
     3361 gaaatttgct tactgtggta aatggtttgt gagtactgga aaagataacc tcctcaatgc
     3421 ttggcggacc ccctatggag ccagcatatt ccagtccaaa gagtcctcgt cagtgcttag
     3481 ctgtgacatc tctgtggatg ataagtacat agtcactggc tcgggggaca agaaggctac
     3541 agtctatgaa gtcatctact aatctagagg tcaagcttcg ggggtttaaa cctgcaggca
     3601 tgcaagctga tttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag
     3661 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3721 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3781 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     3841 aagcaagtaa aacctctaca aatgtggtaa aatcttaatt aagacttcta atataacttc
     3901 gtatagcata cattatacga agttatggtt ccactgttga caattaatca tcggcatagt
     3961 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     4021 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4081 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4141 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4201 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4261 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4321 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4381 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4441 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4501 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4561 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4621 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4681 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4741 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4801 ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     4861 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5401 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5521 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5641 aaaggatctt cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac
     5701 agatgtaatg aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct
     5761 cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac
     5821 tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc
     5881 cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc
     5941 cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa
     6001 aatgatatac attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga
     6061 cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg
     6121 tggattcttc ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct
     6181 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     6241 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     6301 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     6361 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     6421 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     6481 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga
     6541 aggtacctaa ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
