LOCUS       sequence-471875-        4984 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express Histone H4-HaloTag(R) 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 4984)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4984)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Histone H4-HaloTag Fusion Vector
FEATURES             Location/Qualifiers
     source          1..4984
                     /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             1395..1415
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1431..2315
                     /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(2403..2421)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2458..2579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2469..2488)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2523..2542
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             2978..3772
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3032..3051)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3642..3661
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3944..4532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4024..4043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4648..4931
                     /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 cgccatgtcc ggcagaggaa
     1081 agggcggaaa aggcttaggc aaagggggcg ctaagcgcca ccgcaaggtc ttgagagaca
     1141 acattcaggg catcaccaag cctgccattc ggcgtctagc tcggcgtggc ggcgttaagc
     1201 ggatctctgg cctcatttac gaggagaccc gcggtgtgct gaaggtgttc ctggagaatg
     1261 tgattcggga cgcagtcacc tacaccgagc acgccaagcg caagaccgtc acagccatgg
     1321 atgtggtgta cgcgctcaag cgccaggggc gcaccctgta cggcttcgga ggcgtttctc
     1381 tcgagccaac cactgaggat ctgtactttc agagcgataa cgatggatcc gaaatcggta
     1441 ctggctttcc attcgacccc cattatgtgg aagtcctggg cgagcgcatg cactacgtcg
     1501 atgttggtcc gcgcgatggc acccctgtgc tgttcctgca cggtaacccg acctcctcct
     1561 acgtgtggcg caacatcatc ccgcatgttg caccgaccca tcgctgcatt gctccagacc
     1621 tgatcggtat gggcaaatcc gacaaaccag acctgggtta tttcttcgac gaccacgtcc
     1681 gcttcatgga tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg
     1741 actggggctc cgctctgggt ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta
     1801 ttgcatttat ggagttcatc cgccctatcc cgacctggga cgaatggcca gaatttgccc
     1861 gcgagacctt ccaggccttc cgcaccaccg acgtcggccg caagctgatc atcgatcaga
     1921 acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga
     1981 tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc
     2041 caaacgagct gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc gaagaataca
     2101 tggactggct gcaccagtcc cctgtcccga agctgctgtt ctggggcacc ccaggcgttc
     2161 tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg
     2221 acatcggccc gggtctgaat ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga
     2281 tcgcgcgctg gctgtctact ctggagattt ccggttaata gaattctaga gtcgacctgc
     2341 aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2401 caccgctgag caataactag cataacccct tggggcggcc gcttcgagca gacatgataa
     2461 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2521 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2581 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     2641 taagcaagta aaacctctac aaatgtggta aaatcgaatt ctaatggatc ctctttgcgc
     2701 ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt
     2761 tctgcggact ggctttctac gtgttccgct tcctttagca gcccttgcgc cctgagtgct
     2821 tgcggcagcg tgagcttcaa aagaattgcc agctggggcg ccctctggta aggttgggaa
     2881 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
     2941 aagatctgat caagagacag gatgacggtc gtttcgcatg cttgaacaag atggattgca
     3001 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3061 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3121 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3181 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3241 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3301 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3361 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gcactcggat
     3421 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3481 cgaactgttc gccaggctca aggcgcgtat gccggatggt gaggatctcg tcgtgactca
     3541 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3601 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3661 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3721 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3781 ctggggttcg aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg
     3841 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3901 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3961 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4021 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4081 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4141 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4201 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4261 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4321 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4381 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4441 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4501 agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt
     4561 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4621 ggatcttcac ctagatcctt ttatagtccg gaaatacagg aacgcacgct ggatggccct
     4681 tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt
     4741 ggcctgtacc ctctggttgc ataggtattc atacggttaa aatttatcag gcgcgattgc
     4801 ggcagttttt cgggtggttt gttgccattt ttacctgtct gctgccgtga tcgcgctgaa
     4861 cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc
     4921 gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag
     4981 atct
//
