LOCUS       sequence-426001-        8129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human LEO1. N-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8129)
  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 8129)
  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: pHTN-HaloTag_LEO1:1-666
FEATURES             Location/Qualifiers
     source          1..8129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(4080..4098)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4130..4251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4141..4160)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4195..4214
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4496..4853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4496..4687
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4519..4539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4704..4839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4904..5698
                     /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(4958..4977)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5568..5587
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5762..5810
                     /note="synthetic polyadenylation signal"
     promoter        5966..6070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6071..6139,6140..6931)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6289..6308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7089..7677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7169..7188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7793..8076
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgcttccga attcatggcg gatatggagg atctcttcgg
     2041 gagcgacgcc gacagcgaag ctgagcgtaa agattctgat tctggatctg actcagattc
     2101 tgatcaagag aatgctgcct ctggcagtaa tgcctctgga agtgaaagtg atcaggatga
     2161 aagaggtgat tcaggacaac caagtaataa ggaactgttt ggagatgaca gtgaggacga
     2221 gggagcttca catcatagtg gtagtgataa tcactctgaa agatcagaca atagatcaga
     2281 agcttctgag cgttctgacc atgaggacaa tgacccctca gatgtagatc agcacagtgg
     2341 atcagaagcc cctaatgatg atgaagacga aggtcataga tcggatggag ggagccatca
     2401 ttcagaagca gaaggttctg aaaaagcaca ttcagatgat gaaaaatggg gcagagaaga
     2461 taaaagtgac cagtcagatg atgaaaagat acaaaattct gatgatgagg agagggcaca
     2521 aggatctgat gaagataagc tgcagaattc tgacgatgat gagaaaatgc agaacacaga
     2581 tgatgaggag aggcctcagc tttccgatga tgagagacaa cagctatctg aggaggaaaa
     2641 ggctaattct gatgatgaac ggccggtagc ttctgataat gatgatgaga aacagaattc
     2701 tgatgatgaa gaacaaccac agctgtctga tgaagagaaa atgcaaaatt ctgatgatga
     2761 aaggccacag gcctcagatg aagaacacag gcattcagat gatgaagagg aacaggatca
     2821 taaatcagaa tctgcaagag gcagtgatag tgaagatgaa gttttacgaa tgaaacgcaa
     2881 gaatgcgatt gcatctgatt cagaagcgga tagtgacact gaggtgccaa aagataatag
     2941 tggaaccatg gatttatttg gaggtgcaga tgatatctct tcagggagtg atggagaaga
     3001 caaaccacct actccaggac agcctgttga tgaaaatgga ttgcctcagg atcaacagga
     3061 agaggagcca attcctgaga ccagaataga agtagaaata cccaaagtaa acactgattt
     3121 aggaaacgac ttatattttg ttaaactgcc caactttctc agtgtagagc ccagaccttt
     3181 tgatcctcag tattatgaag atgaatttga agatgaagaa atgctggatg aagaaggtag
     3241 aaccaggtta aaattaaagg tagaaaatac tataagatgg aggatacgcc gagatgaaga
     3301 aggaaatgaa attaaagaaa gcaatgctcg gatagtcaag tggtcagatg gaagcatgtc
     3361 cctgcattta ggcaatgaag tgtttgatgt gtacaaagcc ccactgcagg gcgaccacaa
     3421 tcatcttttt ataagacaag gtactggtct acagggacaa gcagtcttta aaacgaaact
     3481 caccttcaga cctcactcta cggacagtgc cacacataga aagatgactc tgtcacttgc
     3541 agataggtgt tcaaagacac agaagattag aatcttgcca atggctggtc gtgatcctga
     3601 atgccaacgc acagaaatga ttaagaaaga agaagaacgt ttgagggctt ccatacgtag
     3661 ggaatctcag cagcgccgaa tgagagagaa acagcaccag cgggggctga gcgccagtta
     3721 cctggaacct gatcgatacg atgaggagga ggaaggcgag gagtccatca gcttggctgc
     3781 cattaaaaac cgatataaag ggggcattcg agaggaacga gccagaatct attcatcaga
     3841 cagtgatgag ggatcagaag aagataaagc tcaaagatta ctcaaagcaa agaaacttac
     3901 cagtgatgag gaaggtgaac cttccggaaa gagaaaagca gaagatgatg ataaagcaaa
     3961 taaaaagcat aagaagtatg tgatcagcga tgaagaggaa gaagatgatg attgatgatc
     4021 tagatttggg cccaattcct gcaggatttt gcggccgctt gctgagttgg ctgctgccac
     4081 cgctgagcaa taactagcat aaccccttgg ccgcttcgag cagacatgat aagatacatt
     4141 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     4201 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     4261 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag
     4321 taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc ttacaatttc
     4381 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct
     4441 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     4501 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4561 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4621 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4681 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4741 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4801 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     4861 attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg
     4921 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4981 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5041 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5101 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5161 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5221 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5281 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5341 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5401 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5461 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5521 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5581 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5641 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5701 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     5761 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     5821 gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     5881 catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt cttagacgtc
     5941 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6001 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6061 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6121 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6181 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6241 ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc tatgtggcgc
     6301 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6361 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6421 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6481 gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6541 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6601 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6661 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6721 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6781 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6841 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6901 gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag cgacgcccaa
     6961 ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7021 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7501 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa
     7681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7741 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaaat
     7801 acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa aatggcagct
     7861 tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg tattcatacg
     7921 gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc catttttacc
     7981 tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta agggattatg
     8041 gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac
     8101 gatgcgtccg gcgtagagga tcgagatct
//
