LOCUS       sequence-470701-        7024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag(R)-Menin 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 7024)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7024)
  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            "Unpublished"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, HaloTag-Menin
FEATURES             Location/Qualifiers
     source          1..7024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3..84
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        317..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          960..1092
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1075..1094
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1136..1155
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1136..1154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1164..1173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1170..2060
                     /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             2073..2093
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     polyA_signal    4066..4187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4077..4096)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4302..4335)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4345..4392
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4424..5218
                     /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(4478..4497)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5088..5107
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5376..5964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5456..5475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6090..6138
                     /note="synthetic polyadenylation signal"
     CDS             complement(6162..6560)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6557..6566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6591..6948)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6605..6740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6757..6948
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6905..6925
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 cggcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
       61 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      121 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
      181 gattatgatc cgcggccgca atacgtcgac gttatcagct gacttcgtac gagagcctag
      241 gattatggcg cgccactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      841 agatcactag aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      901 cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg
      961 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1021 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     1081 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtattaata
     1141 cgactcacta tagggctagc aaagccacca tggcagaaat cggtactggc tttccattcg
     1201 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     1261 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     1321 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     1381 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     1441 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     1501 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     1561 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     1621 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     1681 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     1741 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     1801 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     1861 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     1921 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     1981 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     2041 cgacgctcga gatttccggc gagccaacca ctgaggatct gtactttcag agcgataacg
     2101 cgatcgccat ggggctgaag gccgcccaga agacgctgtt cccgctgcgc tccatcgacg
     2161 acgtggtgcg cctgtttgct gccgagctgg gccgagagga gccggacctg gtgctccttt
     2221 ccttggtgct gggcttcgtg gagcattttc tggctgtcaa ccgcgtcatc cctaccaacg
     2281 ttcccgagct caccttccag cccagccccg cccccgaccc gcctggcggc ctcacctact
     2341 ttcccgtggc cgacctgtct atcatcgccg ccctctatgc ccgcttcacc gcccagatcc
     2401 gaggcgccgt cgacctgtcc ctctatcctc gagaaggggg tgtctccagc cgtgagctgg
     2461 tgaagaaggt ctccgatgtc atatggaaca gcctcagccg ctcctacttc aaggatcggg
     2521 cccacatcca gtccctcttc agcttcatca caggttggag cccagtaggc accaaattgg
     2581 acagctccgg tgtggccttt gctgtggttg gggcctgcca ggccctgggt ctccgggatg
     2641 tccacctcgc cctgtctgag gatcatgcct gggtagtgtt tgggcccaat ggggagcaga
     2701 cagctgaggt cacctggcac ggcaagggca acgaggaccg caggggccag acagtcaatg
     2761 ccggtgtggc tgagcggagc tggctgtacc tgaaaggatc atacatgcgc tgtgaccgca
     2821 agatggaggt ggcgttcatg gtgtgtgcca tcaacccttc cattgacctg cacaccgact
     2881 cgctggagct tctgcagctg cagcagaagc tgctctggct gctctatgac ctgggacatc
     2941 tggaaaggta ccccatggcc ttagggaacc tggcagatct agaggagctg gagcccaccc
     3001 ctggccggcc agacccactc accctctacc acaagggcat tgcctcagcc aagacctact
     3061 atcgggatga acacatctac ccctacatgt acctggctgg ctaccactgt cgcaaccgca
     3121 atgtgcggga agccctgcag gcctgggcgg acacggccac tgtcatccag gactacaact
     3181 actgccggga agacgaggag atctacaagg agttctttga agtagccaat gatgtcatcc
     3241 ccaacctgct gaaggaggca gccagcttgc tggaggcggg cgaggagcgg ccgggggagc
     3301 aaagccaggg cacccagagc caaggttccg ccctccagga ccctgagtgc ttcgcccacc
     3361 tgctgcgatt ctacgacggc atctgcaaat gggaggaggg cagtcccacg cctgtgctgc
     3421 acgtgggctg ggccaccttt cttgtgcagt ccctaggccg ttttgaggga caggtgcggc
     3481 agaaggtgcg catagtgagc cgagaggccg aggcggccga ggccgaggag ccgtggggcg
     3541 aggaagcccg ggaaggccgg cggcggggcc cacggcggga gtccaagcca gaggagcccc
     3601 cgccgcccaa gaagccagca ctggacaagg gcctgggcac cggccagggt gcagtgtcag
     3661 gacccccccg gaagcctcct gggactgtcg ctggcacagc ccgaggccct gaaggtggca
     3721 gcacggctca ggtgccagca cccgcagcat caccaccgcc ggagggtcca gtgctcactt
     3781 tccagagtga gaagatgaag ggcatgaagg agctgctggt ggccaccaag atcaactcga
     3841 gcgccatcaa gctgcaactc acggcacagt cgcaagtgca gatgaagaag cagaaagtgt
     3901 ccacccctag tgactacact ctgtctttcc tcaagcggca gcgcaaaggc ctcgtttaaa
     3961 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     4021 ttggctgctg ccaccgctga gcaaggccgc ttcgagcaga catgataaga tacattgatg
     4081 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4141 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4201 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa
     4261 acctctacaa atgtggtaaa atcttaatta agaattctaa tataacttcg tatagcatac
     4321 attatacgaa gttatggatc cactgttgac aattaatcat cggcatagta tatcggcata
     4381 gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg
     4441 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4501 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4561 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4621 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4681 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4741 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4801 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4861 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4921 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4981 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5041 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5101 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5161 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt
     5221 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     5281 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5341 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5401 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5461 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5521 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5581 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5641 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5701 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5761 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5821 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5881 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5941 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     6001 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6061 acctagatcc ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga
     6121 aaataaagat attttattgc ggccatcgtg atggctagag tttagccctc ccacacataa
     6181 ccagagggca gcaattcacg aatcccaact gccgtcggct gtccatcact gtccttcact
     6241 atggctttga tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc
     6301 aagatgcccc tgttctcatt tccgatcgcg acgatacaag tcaggttgcc agctgccgca
     6361 gcagcagcag tgcccagcac cacgagttct gcacaaggtc ccccagtaaa atgatataca
     6421 ttgacaccag tgaagatgcg gccgtcgcta gagagagctg cgctggcgac gctgtagtct
     6481 tcagagatgg ggatgctgtt gattgtagcc gttgctcttt caatgagggt ggattcttct
     6541 tgagacaaag gcttggccat ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag
     6601 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     6661 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     6721 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     6781 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     6841 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     6901 ccacacccta actgacacac attccacagc tggttctttc cgcctcagaa ggtacctaac
     6961 caagttcctc tttcagaggt tatttcaggc catggtgctg cgcaagacgc gttaattaag
     7021 acgt
//
