LOCUS       sequence-424689-        6638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human TLE1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6638)
  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 6638)
  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_TLE1:1-770
FEATURES             Location/Qualifiers
     source          1..6638
                     /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    3701..3822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3712..3731)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3937..3970)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3980..4027
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4059..4853
                     /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(4113..4132)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4723..4742
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5011..5599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5091..5110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5725..5773
                     /note="synthetic polyadenylation signal"
     CDS             complement(5797..6195)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6192..6201
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6226..6583)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6240..6375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6392..6583
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6540..6560
                     /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 ccatgttccc gcagagccgg
     1261 cacccgacgc cgcaccaggc tgcaggccag cccttcaagt tcactatccc ggagtccctg
     1321 gaccggatta aagaggaatt ccagttcctg caggcgcagt atcacagcct taaattggaa
     1381 tgtgagaaac tggcaagtga aaagacagaa atgcagaggc actatgtgat gtattatgaa
     1441 atgtcatatg gattaaacat tgaaatgcac aaacagactg aaatcgccaa gagattgaat
     1501 acgatttgtg cacaagtcat cccatttctg tctcaggaac atcaacaaca ggtggcccag
     1561 gctgttgaac gtgccaaaca ggtgaccatg gcagagttga atgccatcat cgggcagcag
     1621 cagttgcaag ctcagcatct ttctcatggc cacggacccc cagttcccct tacgcctcac
     1681 ccttcgggac ttcagcctcc tggaatcccg cccctcgggg gcagtgccgg ccttcttgcg
     1741 ctgtctagtg ctctgagtgg gcagtctcac ttggcaataa aagatgacaa gaagcaccac
     1801 gatgcagagc accacagaga cagagagccg ggcacaagta attccctcct ggtcccagac
     1861 agtctaagag gcacagataa acgcagaaat ggacctgaat tttccaatga catcaagaaa
     1921 aggaaggtgg atgataagga ctccagccac tatgacagtg atggtgacaa aagcgatgac
     1981 aacttagttg tggatgtgtc taatgaggac ccttcttctc cgcgagcaag ccctgcccac
     2041 tcgccccggg aaaatggaat cgacaaaaat cgcctgctaa agaaggatgc ttctagcagt
     2101 ccagcttcca cggcctcctc ggcaagttcc acttctttga aatccaaaga aatgagcttg
     2161 catgaaaaag ccagcacgcc tgttctgaaa tccagcacac caacgcctcg gagcgacatg
     2221 ccaacgccgg gcaccagcgc cactccaggc ctccgtccag gtctcggcaa gcctccagcc
     2281 atagaccccc tcgttaacca agcggcagct ggcttgagga cacccctggc agtgcccggc
     2341 ccatatcctg ctccttttgg gatggtcccc cacgctggca tgaacggcga gctgaccagc
     2401 ccaggcgctg cctacgccag tttacacaac atgtcgcccc agatgagcgc cgcagccgcc
     2461 gcggccgccg tggtggccta cgggcgctcc cccatggtgg ggtttgatcc tccccctcac
     2521 atgagagtac ctaccattcc tccaaacctg gcaggaatcc ctggggggaa acctgcatac
     2581 tccttccacg ttactgcaga cggtcagatg cagcctgtcc cctttccccc cgacgccctc
     2641 atcggacccg gaatcccccg gcatgctcgc cagatcaaca ccctcaacca cggggaggtg
     2701 gtgtgcgctg tgaccatcag caaccccacg agacacgtgt acacaggcgg gaagggctgc
     2761 gtcaaggtct gggacatcag ccaccctggc aataagagcc ctgtctccca gctcgactgt
     2821 ctgaacagag acaattatat ccgttcctgt aaattgctac ccgatggctg cactctcata
     2881 gtgggagggg aagccagtac tttgtccatt tgggacctgg cggctccaac cccgcgcatc
     2941 aaggcggagc tgacgtcctc ggcccccgcc tgctacgccc tggccatcag ccccgattcc
     3001 aaggtctgct tctcatgctg cagcgacggc aacatcgctg tgtgggatct gcacaaccag
     3061 acactagtga ggcaattcca gggccacaca gacggagcca gctgtattga catttctaat
     3121 gatggcacca agctctggac gggtggtttg gacaacacag tcaggtcctg ggacctgcgc
     3181 gaggggcggc agctgcagca gcacgacttc acctcccaga tcttctccct ggggtactgc
     3241 cccaccgggg agtggctggc agtgggcatg gagagcagca atgtggaggt gctgcacgtg
     3301 aacaagcctg acaagtacca gctgcacctg catgagagct gcgtgctgtc cctgaaattt
     3361 gcttactgtg gtaaatggtt tgtgagtact ggaaaagata acctcctcaa tgcttggcgg
     3421 accccctatg gagccagcat attccagtcc aaagagtcct cgtcagtgct tagctgtgac
     3481 atctctgtgg atgataagta catagtcact ggctcggggg acaagaaggc tacagtctat
     3541 gaagtcatct acggctcgag cggtgggagc tccggtgtga gcggctggcg gctgttcaag
     3601 aagattagct aagtttaaac ggccgcgact ctagactgca ggcatgcaag ctgatttggc
     3661 tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat tgatgagttt
     3721 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3781 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3841 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     3901 tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag catacattat
     3961 acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg gcatagtata
     4021 atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc
     4081 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4141 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4201 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4261 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4321 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4381 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4441 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4501 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4561 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4621 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4681 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4741 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4801 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaaa
     4861 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     4921 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4981 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5041 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5101 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5161 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5221 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5281 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5341 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5401 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5461 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5521 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5581 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5641 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5701 gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt aatgaaaata
     5761 aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca cataaccaga
     5821 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     5881 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     5941 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     6001 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     6061 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     6121 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     6181 caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag
     6241 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     6301 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     6361 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     6421 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     6481 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6541 ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac ctaaccaagt
     6601 tcctctttca gaggttattt caggccatgg tgctgcgc
//
