LOCUS       sequence-474934-        7173 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 7173)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7173)
  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            "Unpublished"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..7173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1326..1346
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1362..2246
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    2333..2454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2344..2363)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2569..2602)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2612..2659
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2691..3485
                     /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(2745..2764)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3355..3374
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3702..4247
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4148..4167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4364..4381
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4531..4579
                     /note="synthetic polyadenylation signal"
     CDS             complement(4603..5001)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4998..5007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5032..5389)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5046..5181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5198..5389
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5346..5366
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    5468..5549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        5782..6085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6086..6289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6240..6260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          6425..6557
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     6540..6559
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     6601..6620
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6601..6619
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccctgtctga ggatcatgcc tgggtagtgt ttgggcccaa tggggagcag acagctgagg
       61 tcacctggca cggcaagggc aacgaggacc gcaggggcca gacagtcaat gccggtgtgg
      121 ctgagcggag ctggctgtac ctgaaaggat catacatgcg ctgtgaccgc aagatggagg
      181 tggcgttcat ggtgtgtgcc atcaaccctt ccattgacct gcacaccgac tcgctggagc
      241 ttctgcagct gcagcagaag ctgctctggc tgctctatga cctgggacat ctggaaaggt
      301 accccatggc cttagggaac ctggcagatc tagaggagct ggagcccacc cctggccggc
      361 cagacccact caccctctac cacaagggca ttgcctcagc caagacctac tatcgggatg
      421 aacacatcta cccctacatg tacctggctg gctaccactg tcgcaaccgc aatgtgcggg
      481 aagccctgca ggcctgggcg gacacggcca ctgtcatcca ggactacaac tactgccggg
      541 aagacgagga gatctacaag gagttctttg aagtagccaa tgatgtcatc cccaacctgc
      601 tgaaggaggc agccagcttg ctggaggcgg gcgaggagcg gccgggggag caaagccagg
      661 gcacccagag ccaaggttcc gccctccagg accctgagtg cttcgcccac ctgctgcgat
      721 tctacgacgg catctgcaaa tgggaggagg gcagtcccac gcctgtgctg cacgtgggct
      781 gggccacctt tcttgtgcag tccctaggcc gttttgaggg acaggtgcgg cagaaggtgc
      841 gcatagtgag ccgagaggcc gaggcggccg aggccgagga gccgtggggc gaggaagccc
      901 gggaaggccg gcggcggggc ccacggcggg agtccaagcc agaggagccc ccgccgccca
      961 agaagccagc actggacaag ggcctgggca ccggccaggg tgcagtgtca ggaccccccc
     1021 ggaagcctcc tgggactgtc gctggcacag cccgaggccc tgaaggtggc agcacggctc
     1081 aggtgccagc acccgcagca tcaccaccgc cggagggtcc agtgctcact ttccagagtg
     1141 agaagatgaa gggcatgaag gagctgctgg tggccaccaa gatcaactcg agcgccatca
     1201 agctgcaact cacggcacag tcgcaagtgc agatgaagaa gcagaaagtg tccaccccta
     1261 gtgactacac tctgtctttc ctcaagcggc agcgcaaagg cctcgtttct ctcgagccaa
     1321 ccactgagga tctgtacttt cagagcgata acgatggatc cgaaatcggt actggctttc
     1381 cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc
     1441 cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc
     1501 gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta
     1561 tgggcaaatc cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg
     1621 atgccttcat cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct
     1681 ccgctctggg tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta
     1741 tggagttcat ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct
     1801 tccaggcctt ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta
     1861 tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt
     1921 accgcgagcc gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc
     1981 tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc
     2041 tgcaccagtc ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac
     2101 cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc
     2161 cgggtctgaa tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct
     2221 ggctgtctac tctggagatt tccggttaat agaattctag agtcgacctg caggcatgca
     2281 agctgatttg gctgctgcca ccgctgagca aggccgcttc gagcagacat gataagatac
     2341 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2401 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2461 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     2521 aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga attctaatat aacttcgtat
     2581 agcatacatt atacgaagtt atggatccac tgttgacaat taatcatcgg catagtatat
     2641 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     2701 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2761 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2821 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2881 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2941 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3001 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3061 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3121 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3181 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3241 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3301 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3361 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3421 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3481 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     3541 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtcaa acatgagaat
     3601 tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
     3661 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
     3721 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
     3781 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
     3841 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     3901 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     3961 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     4021 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     4081 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     4141 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     4201 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     4261 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     4321 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     4381 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     4441 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     4501 cactccgcta gcgctgatgt ccggaatcgt cacacaaaaa accaacacac agatgtaatg
     4561 aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct cccacacata
     4621 accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac
     4681 tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc cgcaggggct
     4741 caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc
     4801 agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa aatgatatac
     4861 attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc
     4921 ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc
     4981 ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa
     5041 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     5101 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     5161 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     5221 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     5281 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5341 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
     5401 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgcaagacg cgttaattaa
     5461 gacgtcggca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5521 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     5581 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     5641 tggctgatta tgatccgcgg ccgcaatacg tcgacgttat cagctgactt cgtacgagag
     5701 cctaggatta tggcgcgcca ctagttatta atagtaatca attacggggt cattagttca
     5761 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5821 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5881 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5941 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     6001 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     6061 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     6121 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6181 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     6241 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     6301 ccgtcagatc actagaagct ttattgcggt agtttatcac agttaaattg ctaacgcagt
     6361 cagtgcttct gacacaacag tctcgaactt aagctgcaga agttggtcgt gaggcactgg
     6421 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
     6481 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
     6541 tgcctttctc tccacaggtg tccactccca gttcaattac agctcttaag gctagagtat
     6601 taatacgact cactataggg ctagcgatcg ccatggggct gaaggccgcc cagaagacgc
     6661 tgttcccgct gcgctccatc gacgacgtgg tgcgcctgtt tgctgccgag ctgggccgag
     6721 aggagccgga cctggtgctc ctttccttgg tgctgggctt cgtggagcat tttctggctg
     6781 tcaaccgcgt catccctacc aacgttcccg agctcacctt ccagcccagc cccgcccccg
     6841 acccgcctgg cggcctcacc tactttcccg tggccgacct gtctatcatc gccgccctct
     6901 atgcccgctt caccgcccag atccgaggcg ccgtcgacct gtccctctat cctcgagaag
     6961 ggggtgtctc cagccgtgag ctggtgaaga aggtctccga tgtcatatgg aacagcctca
     7021 gccgctccta cttcaaggat cgggcccaca tccagtccct cttcagcttc atcacaggtt
     7081 ggagcccagt aggcaccaaa ttggacagct ccggtgtggc ctttgctgtg gttggggcct
     7141 gccaggccct gggtctccgg gatgtccacc tcg
//
