LOCUS       sequence-473372-        5707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag(R)-WDR5 Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5707)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5707)
  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: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay Vector, WDR5
FEATURES             Location/Qualifiers
     source          1..5707
                     /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(3121..3139)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3176..3297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3187..3206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3241..3260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3544..3648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3649..3717,3718..4509)
                     /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(3867..3886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4667..5255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4747..4766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5371..5654
                     /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 tcgccatggc gacggaggag aagaagcccg agaccgaggc
     2041 cgccagagca cagccaaccc cttcgtcatc cgccactcag agcaagccta cacctgtgaa
     2101 gccaaactat gctctaaagt tcacccttgc tggccacacc aaagcagtgt cctccgtgaa
     2161 attcagcccg aatggagagt ggctggcaag ttcatctgct gataaactta ttaaaatttg
     2221 gggcgcgtat gatgggaaat ttgagaaaac catatctggt cacaagctgg gaatatccga
     2281 tgtagcctgg tcgtcagatt ctaaccttct tgtttctgcc tcagatgaca aaaccttgaa
     2341 gatatgggac gtgagctcgg gcaagtgtct gaaaaccctg aagggacaca gtaattatgt
     2401 cttttgctgc aacttcaatc cccagtccaa ccttattgtc tcaggatcct ttgacgaaag
     2461 cgtgaggata tgggatgtga aaacagggaa gtgcctcaag actttgccag ctcactcgga
     2521 tccagtctcg gccgttcatt ttaatcgtga tggatccttg atagtttcaa gtagctatga
     2581 tggtctctgt cgcatctggg acaccgcctc aggccagtgc ctgaagacgc tcatcgatga
     2641 cgacaacccc cccgtgtctt ttgtgaagtt ctccccgaac ggcaaataca tcctggccgc
     2701 cacgctggac aacactctga agctctggga ctacagcaag gggaagtgcc tgaagacgta
     2761 cactggccac aagaatgaga aatactgcat atttgccaat ttctctgtta ctggtgggaa
     2821 gtggattgtg tctggctcag aggataacct tgtttacatc tggaaccttc agacgaaaga
     2881 gattgtacag aaactacaag gccacacaga tgtcgtgatc tcaacagctt gtcacccaac
     2941 agaaaacatc atcgcctctg ctgcgctaga aaatgacaaa acaattaaac tgtggaagag
     3001 tgactgcgtt taaacgaatt cgggctcggt acccggggat cctctagagt cgacctgcag
     3061 gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     3121 ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga
     3181 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3241 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3301 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta
     3361 agcaagtaaa acctctacaa atgtggtaaa atcgaattct aatggatcct ctttgcgctt
     3421 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     3481 tgcggactgg ctttctacgt aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3541 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3601 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3661 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3721 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3781 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3841 ccaatgatga gcactttcaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3901 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3961 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4021 ataaccatga gtgataacac tgcggccaac ttacttctga caactatcgg aggaccgaag
     4081 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4141 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4201 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4261 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4321 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4381 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4441 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4501 cattggtaat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     4561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5101 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5221 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5341 aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc
     5401 ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa
     5461 tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat
     5521 tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct
     5581 gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     5641 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     5701 gagatct
//
