LOCUS       sequence-479503-        6354 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Express BRD4(46-460)-HaloTag 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 6354)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6354)
  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            "Unpublished"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: BRD4(46-460)-HaloTag Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6354
                     /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"
     CDS             2289..2309
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2325..3209
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(3297..3315)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3352..3473
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3363..3382)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3417..3436
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3718..4075
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3718..3909
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3741..3761)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3926..4061
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4090..4137
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4169..4963
                     /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(4223..4242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4833..4852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5027..5075
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5314..5902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5394..5413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6018..6301
                     /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 ggctagcgat cgccatggag acctccaacc
     1081 ctaacaagcc caagaggcag accaaccaac tgcaatacct gctcagagtg gtgctcaaga
     1141 cactatggaa acaccagttt gcatggcctt tccagcagcc tgtggatgcc gtcaagctga
     1201 acctccctga ttactataag atcattaaaa cgcctatgga tatgggaaca ataaagaagc
     1261 gcttggaaaa caactattac tggaatgctc aggaatgtat ccaggacttc aacactatgt
     1321 ttacaaattg ttacatctac aacaagcctg gagatgacat agtcttaatg gcagaagctc
     1381 tggaaaagct cttcttgcaa aaaataaatg agctacccac agaagaaacc gagatcatga
     1441 tagtccaggc aaaaggaaga ggacgtggga ggaaagaaac agggacagca aaacctggcg
     1501 tttccacggt accaaacaca actcaagcat cgactcctcc gcagacccag acccctcagc
     1561 cgaatcctcc tcctgtgcag gccacgcctc accccttccc tgccgtcacc ccggacctca
     1621 tcgtccagac ccctgtcatg acagtggtgc ctccccagcc actgcagacs cccccgccag
     1681 tgccccccca gccacaaccc ccacccgctc cagctcccca gcccgtacag agccacccac
     1741 ccatcatcgc ggccacccca cagcctgtga agacaaagac caccattgac cccattcacg
     1801 agccaccctc gctgcccccg gagcccaaga ccaccaagct gggccagcgg cgggagagca
     1861 gccggcctgt gaaacctcca aagaaggacg tgcccgactc tcagcagcac ccagcaccag
     1921 agaagagcag caaggtctcg gagcagctca agtgctgcag cggcatcctc aaggagatgt
     1981 ttgccaagaa gcacgccgcc tacgcctggc ccttctacaa gcctgtggac gtggaggcac
     2041 tgggcctaca cgactactgt gacatcatca agcaccccat ggacatgagc acaatcaagt
     2101 ctaaactgga ggcccgtgag taccgtgatg ctcaggagtt tggtgctgac gtccgattga
     2161 tgttctccaa ctgctataag tacaaccctc ctgaccatga ggtggtggcc atggcccgca
     2221 agctccagga tgtgttcgaa atgcgctttg ccaagatgcc ggacgaggtt tctctcgagc
     2281 caaccactga ggatctgtac tttcagagcg ataacgatgg atccgaaatc ggtactggct
     2341 ttccattcga cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg
     2401 gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt
     2461 ggcgcaacat catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg
     2521 gtatgggcaa atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca
     2581 tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg
     2641 gctccgctct gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat
     2701 ttatggagtt catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga
     2761 ccttccaggc cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt
     2821 ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc
     2881 attaccgcga gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg
     2941 agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact
     3001 ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc
     3061 caccggccga agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg
     3121 gcccgggtct gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc
     3181 gctggctgtc tactctggag atttccggtt aatagaattc tagagtcgac ctgcaggcat
     3241 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     3301 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     3361 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3421 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3481 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     3541 agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt
     3601 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     3661 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     3721 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3781 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3841 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3901 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3961 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4021 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4081 taattaactg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     4141 gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc
     4201 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4261 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4321 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4381 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4441 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4501 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4561 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4621 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4681 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4741 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4801 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4861 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4921 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4981 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     5041 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatcctctt
     5101 tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca
     5161 ccgtttctgc ggactggctt tctacccggt atcagctcac tcaaaggcgg taatacggtt
     5221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5881 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6001 ctagatcctt ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat
     6061 ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc
     6121 ctctggttgc ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt
     6181 cgggtggttt gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag
     6241 cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg
     6301 agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
