LOCUS       sequence-393694-        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDR5 with C-terminal HaloTag fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  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 6001)
  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: WDR5-HaloTag
FEATURES             Location/Qualifiers
     source          1..6001
                     /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             3096..3983
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     primer_bind     complement(4094..4112)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4149..4270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4160..4179)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4214..4233
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4515..4872
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4515..4706
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4538..4558)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4723..4858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4887..4934
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4966..5760
                     /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(5020..5039)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5630..5649
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5824..5872
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg
       61 ctctgaaaac ggaaaaaccg 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 ggtcgcgacg gaggagaaga
     2101 agcccgagac cgaggccgcc agagcacagc caaccccttc gtcatccgcc actcagagca
     2161 agcctacacc tgtgaagcca aactatgctc taaagttcac ccttgctggc cacaccaaag
     2221 cagtgtcctc cgtgaaattc agcccgaatg gagagtggct ggcaagttca tctgctgata
     2281 aacttattaa aatttggggc gcgtatgatg ggaaatttga gaaaaccata tctggtcaca
     2341 agctgggaat atccgatgta gcctggtcgt cagattctaa ccttcttgtt tctgcctcag
     2401 atgacaaaac cttgaagata tgggacgtga gctcgggcaa gtgtctgaaa accctgaagg
     2461 gacacagtaa ttatgtcttt tgctgcaact tcaatcccca gtccaacctt attgtctcag
     2521 gatcctttga cgaaagcgtg aggatatggg atgtgaaaac agggaagtgc ctcaagactt
     2581 tgccagctca ctcggatcca gtctcggccg ttcattttaa tcgtgatgga tccttgatag
     2641 tttcaagtag ctatgatggt ctctgtcgca tctgggacac cgcctcgggc cagtgcctga
     2701 agacgctcat cgatgacgac aacccccccg tgtcttttgt gaagttctcc ccgaacggca
     2761 aatacatcct ggccgccacg ctggacaaca ctctgaagct ctgggactac agcaagggga
     2821 agtgcctgaa gacgtacact ggccacaaga atgagaaata ctgcatattt gccaatttct
     2881 ctgttactgg tgggaagtgg attgtgtctg gctcagagga taaccttgtt tacatctgga
     2941 accttcagac gaaagagatt gtacagaaac tacaaggcca cacagatgtc gtgatctcaa
     3001 cagcttgtca cccaacagaa aacatcatcg cctctgctgc gctagaaaat gacaaaacaa
     3061 ttaaactgtg gaagagtgac tgcggatcgt cgggagcaga aatcggtact ggctttccat
     3121 tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc
     3181 gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca
     3241 acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg
     3301 gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg
     3361 ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg
     3421 ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg
     3481 agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc
     3541 aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg
     3601 agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc
     3661 gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc
     3721 caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc
     3781 accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg
     3841 ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg
     3901 gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc
     3961 tgtcgacgct cgagatttcc ggctaaacga attcgggctc ggtacccggg gatcctctag
     4021 agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag
     4081 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc
     4141 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4201 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4261 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     4321 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa
     4381 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4441 acaccgcata cgcggatctg cgcaacacca tggcctgaaa taacctctga aagaggaact
     4501 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4561 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4621 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4681 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4741 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4801 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4861 ggcttttgca aaaagcttaa ttaactgttg acaattaatc atcggcatag tatatcggca
     4921 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     4981 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5041 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5101 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5161 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5221 gaagcgggaa gggactggct gctattgggc gaagtgccgg gacaggatct cctgtcatct
     5281 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5341 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5401 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5461 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5521 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5581 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5641 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5701 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5761 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     5821 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     5881 gcgataagga tccgctcatg cgatgataag ctgtcaaaca tgagaattac aacttatatc
     5941 gtatggggct gacttcaggt gctacatttg aagagataaa ttgcactgaa atctagaaat
     6001 a
//
