LOCUS       sequence-479501-        6379 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Express HaloTag-BRD4(46-460) 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 6379)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6379)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HaloTag-BRD4(46-460) Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6379
                     /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(3322..3340)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3377..3498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3388..3407)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3442..3461
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3743..4100
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3743..3934
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3766..3786)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3951..4086
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4115..4162
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4194..4988
                     /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(4248..4267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4858..4877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5052..5100
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5339..5927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5419..5438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6043..6326
                     /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 tcgccatgga gacctccaac cctaacaagc ccaagaggca
     2041 gaccaaccaa ctgcaatacc tgctcagagt ggtgctcaag acactatgga aacaccagtt
     2101 tgcatggcct ttccagcagc ctgtggatgc cgtcaagctg aacctccctg attactataa
     2161 gatcattaaa acgcctatgg atatgggaac aataaagaag cgcttggaaa acaactatta
     2221 ctggaatgct caggaatgta tccaggactt caacactatg tttacaaatt gttacatcta
     2281 caacaagcct ggagatgaca tagtcttaat ggcagaagct ctggaaaagc tcttcttgca
     2341 aaaaataaat gagctaccca cagaagaaac cgagatcatg atagtccagg caaaaggaag
     2401 aggacgtggg aggaaagaaa cagggacagc aaaacctggc gtttccacgg taccaaacac
     2461 aactcaagca tcgactcctc cgcagaccca gacccctcag ccgaatcctc ctcctgtgca
     2521 ggccacgcct caccccttcc ctgccgtcac cccggacctc atcgtccaga cccctgtcat
     2581 gacagtggtg cctccccagc cmctgcagac scccccgcca gtgccccccc agccacaacc
     2641 cccacccgct ccagctcccc agcccgtaca gagccaccca cccatcatcg cggccacccc
     2701 acagcctgtg aagacaaaga ccaccattga ccccattcac gagccaccct cgctgccccc
     2761 ggagcccaag accaccaagc tgggccagcg gcgggagagc agccggcctg tgaaacctcc
     2821 aaagaaggac gtgcccgact ctcagcagca cccagcacca gagaagagca gcaaggtctc
     2881 ggagcagctc aagtgctgca gcggcatcct caaggagatg tttgccaaga agcacgccgc
     2941 ctacgcctgg cccttctaca agcctgtgga cgtggaggca ctgggcctac acgactactg
     3001 tgacatcatc aagcacccca tggacatgag cacaatcaag tctaaactgg aggcccgtga
     3061 gtaccgtgat gctcaggagt ttggtgctga cgtccgattg atgttctcca actgctataa
     3121 gtacaaccct cctgaccatg aggtggtggc catggcccgc aagctccagg atgtgttcga
     3181 aatgcgcttt gccaagatgc cggacgaggt ttaaacgaat tcgggctcgg tacccgggga
     3241 tcctctagag tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg
     3301 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcggccg
     3361 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3421 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3481 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3541 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcgaattt
     3601 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     3661 ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa
     3721 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3781 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3841 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3901 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3961 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4021 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4081 gaggcctagg cttttgcaaa aagcttaatt aactgttgac aattaatcat cggcatagta
     4141 tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgattg
     4201 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4261 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4321 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4381 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4441 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4501 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4561 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4621 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4681 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4741 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4801 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4861 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4921 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4981 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5041 acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     5101 aatcgatagc gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     5161 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac ccggtatcag
     5221 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5281 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5341 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5401 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5461 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5521 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5581 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5641 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5701 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5761 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5821 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5881 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga
     5941 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6001 tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaaat acaggaacgc
     6061 acgctggatg gcccttcgct gggatggtga aaccatgaaa aatggcagct tcagtggatt
     6121 aagtgggggt aatgtggcct gtaccctctg gttgcatagg tattcatacg gttaaaattt
     6181 atcaggcgcg attgcggcag tttttcgggt ggtttgttgc catttttacc tgtctgctgc
     6241 cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta agggattatg gtaaatccac
     6301 ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac gatgcgtccg
     6361 gcgtagagga tcgagatct
//
