LOCUS       sequence-393695-        6004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDR5 with N-terminal HaloTag fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6004)
  AUTHORS   Schwalm MP, Kramer A, Dolle A, Weckesser J, Yu X, Jin J, Saxena K, 
            Knapp S
  TITLE     Tracking the PROTAC degradation pathway in living cells highlights 
            the importance of ternary complex measurement for PROTAC 
            optimization.
  JOURNAL   Cell Chem Biol. 2023 Jun 20:S2451-9456(23)00157-5. doi: 
            10.1016/j.chembiol.2023.06.002.
  PUBMED    37354907
REFERENCE   2  (bases 1 to 6004)
  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"; journalName: "Cell Chem 
            Biol. 2023 Jun 20:S2451-9456(23)00157-5. doi: 
            10.1016/j.chembiol.2023.06.002."
COMMENT     Sequence Label: HaloTag-WDR5
FEATURES             Location/Qualifiers
     source          1..6004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..573
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     474..493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     690..707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        1152..1531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1532..1735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1686..1706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1871..2003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1986..2005
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     2047..2066
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2047..2065
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2079..2972
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MVAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTS
                     SYVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLV
                     IHDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLI
                     IDQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVAL
                     VEEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPD
                     LIGSEIARWLSTLEISG"
     primer_bind     complement(4097..4115)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4152..4273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4163..4182)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4217..4236
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4518..4875
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4518..4709
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4541..4561)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4726..4861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4890..4937
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4969..5763
                     /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(5023..5042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5633..5652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5827..5875
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg
       61 ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa
      121 ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt
      181 ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg
      241 ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg
      361 cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg
      421 acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac
      481 gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg
      541 tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt
      601 ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt
      661 ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata
      721 tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag
      781 cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc
      841 tgatgtccgg cggtgctttt gccgttacgc accaccccgt catgagatta cgcgttttag
      901 cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg
      961 agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag atcttcaata
     1021 ttggccatta gccatattat tcattggtta tatagcataa atcaatattg gctattggcc
     1081 attgcatacg ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg
     1141 accgccatgt tggcattgat tattgactag ttattaatag taatcaatta cggggtcatt
     1201 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     1261 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     1321 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     1381 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
     1441 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
     1501 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
     1561 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     1621 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
     1681 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     1741 agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt aaattgctaa
     1801 cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt ggtcgtgagg
     1861 cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     1921 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
     1981 ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct cttaaggcta
     2041 gagtattaat acgactcact atagggctag cgctcaccat ggtcgcagaa atcggtactg
     2101 gctttccatt cgacccccat tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg
     2161 ttggtccgcg cgatggcacc cctgtgctgt tcctgcacgg taacccgacc tcctcctacg
     2221 tgtggcgcaa catcatcccg catgttgcac cgacccatcg ctgcattgct ccagacctga
     2281 tcggtatggg caaatccgac aaaccagacc tgggttattt cttcgacgac cacgtccgct
     2341 tcatggatgc cttcatcgaa gccctgggtc tggaagaggt cgtcctggtc attcacgact
     2401 ggggctccgc tctgggtttc cactgggcca agcgcaatcc agagcgcgtc aaaggtattg
     2461 catttatgga gttcatccgc cctatcccga cctgggacga atggccagaa tttgcccgcg
     2521 agaccttcca ggccttccgc accaccgacg tcggccgcaa gctgatcatc gatcagaacg
     2581 tttttatcga gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg
     2641 accattaccg cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa
     2701 acgagctgcc aatcgccggt gagccagcga acatcgtcgc gctggtcgaa gaatacatgg
     2761 actggctgca ccagtcccct gtcccgaagc tgctgttctg gggcacccca ggcgttctga
     2821 tcccaccggc cgaagccgct cgcctggcca aaagcctgcc taactgcaag gctgtggaca
     2881 tcggcccggg tctgaatctg ctgcaagaag acaacccgga cctgatcggc agcgagatcg
     2941 cgcgctggct gtcgacgctc gagatttccg gcggatcgtc gggaatggcg acggaggaga
     3001 agaagcccga gaccgaggcc gccagagcac agccaacccc ttcgtcatcc gccactcaga
     3061 gcaagcctac acctgtgaag ccaaactatg ctctaaagtt cacccttgct ggccacacca
     3121 aagcagtgtc ctccgtgaaa ttcagcccga atggagagtg gctggcaagt tcatctgctg
     3181 ataaacttat taaaatttgg ggcgcgtatg atgggaaatt tgagaaaacc atatctggtc
     3241 acaagctggg aatatccgat gtagcctggt cgtcagattc taaccttctt gtttctgcct
     3301 cagatgacaa aaccttgaag atatgggacg tgagctcggg caagtgtctg aaaaccctga
     3361 agggacacag taattatgtc ttttgctgca acttcaatcc ccagtccaac cttattgtct
     3421 caggatcctt tgacgaaagc gtgaggatat gggatgtgaa aacagggaag tgcctcaaga
     3481 ctttgccagc tcactcggat ccagtctcgg ccgttcattt taatcgtgat ggatccttga
     3541 tagtttcaag tagctatgat ggtctctgtc gcatctggga caccgcctcg ggccagtgcc
     3601 tgaagacgct catcgatgac gacaaccccc ccgtgtcttt tgtgaagttc tccccgaacg
     3661 gcaaatacat cctggccgcc acgctggaca acactctgaa gctctgggac tacagcaagg
     3721 ggaagtgcct gaagacgtac actggccaca agaatgagaa atactgcata tttgccaatt
     3781 tctctgttac tggtgggaag tggattgtgt ctggctcaga ggataacctt gtttacatct
     3841 ggaaccttca gacgaaagag attgtacaga aactacaagg ccacacagat gtcgtgatct
     3901 caacagcttg tcacccaaca gaaaacatca tcgcctctgc tgcgctagaa aatgacaaaa
     3961 caattaaact gtggaagagt gactgctaaa cgaattcggg ctcggtaccc ggggatcctc
     4021 tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct
     4081 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg
     4141 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4201 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4261 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     4321 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     4381 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4441 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     4501 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4681 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4741 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4801 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4861 ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca tagtatatcg
     4921 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     4981 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5041 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5101 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5161 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5221 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cgggacagga tctcctgtca
     5281 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5341 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5401 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5461 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5521 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5581 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5641 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5701 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5761 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     5821 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     5881 atagcgataa ggatccgctc atgcgatgat aagctgtcaa acatgagaat tacaacttat
     5941 atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga
     6001 aata
//
