LOCUS       sequence-475171-        6637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag-PPP2R5A 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 6637)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6637)
  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            "Unpublished"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-PPP2R5A
FEATURES             Location/Qualifiers
     source          1..6637
                     /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    3679..3800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3690..3709)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3915..3948)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3958..4005
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4037..4831
                     /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(4091..4110)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4701..4720
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4989..5577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5069..5088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5703..5751
                     /note="synthetic polyadenylation signal"
     CDS             complement(5775..6173)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6170..6179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6204..6561)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6218..6353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6370..6561
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6518..6538
                     /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 gtcgtcgtcg tcgccgccgg cgggggctgc cagcgccgcc atctcggcct
     2161 cggagaaagt ggacggcttc acccggaaat cggtccgcaa ggcgcagagg cagaagcgct
     2221 cccagggctc gtcgcagttt cgcagccagg gcagccaggc agagctgcac ccgctgcccc
     2281 agctcaaaga tgccacttca aatgaacaac aagagctttt ctgtcagaag ttgcagcagt
     2341 gttgtatact gtttgatttc atggactctg tttcagactt gaagagcaaa gaaattaaaa
     2401 gagcaacact gaatgaactg gttgagtatg tttcaactaa tcgtggtgta attgttgaat
     2461 cagcgtattc tgatatagta aaaatgatca gtgctaacat cttccgtaca cttcctccaa
     2521 gtgataatcc agattttgat ccagaagagg atgaacccac gcttgaggcc tcttggcctc
     2581 acatacagtt ggtatatgaa ttcttcttga gatttttgga gagccctgat ttccagccta
     2641 gcattgcaaa acgatacatt gatcagaaat tcgtacaaca gctcctggag ctttttgata
     2701 gtgaagatcc cagagaacgt gacttcctga agactgttct gcaccgaatt tatgggaaat
     2761 ttcttggatt aagagcattc atcagaaaac aaattaacaa cattttcctc aggtttatat
     2821 atgaaacaga acatttcaat ggtgttgctg aacttcttga aatattagga agtattatca
     2881 atggctttgc attgccactg aaagcagaac ataaacaatt tctaatgaag gttcttattc
     2941 ctatgcatac tgcaaaagga ttagctttgt ttcatgctca gctagcatat tgtgttgtac
     3001 agttcctgga gaaagataca acactaacag agccagtgat cagaggactg ctgaaatttt
     3061 ggccaaaaac ctgcagtcag aaagaggtga tgtttttagg agaaattgaa gaaatcttag
     3121 atgtcattga accaacacag ttcaaaaaaa ttgaagagcc acttttcaag cagatatcca
     3181 agtgtgtatc cagttctcat tttcaggttg cagaaagggc attgtacttc tggaataacg
     3241 aatatattct tagtttgatt gaggagaaca ttgataaaat tctgccaatt atgtttgcca
     3301 gtttgtacaa aatttccaaa gaacactgga atccgaccat tgtagcactg gtatacaatg
     3361 tgctgaaaac cctaatggaa atgaatggca agcttttcga tgaccttact agctcataca
     3421 aagctgaaag acagagagag aaaaagaagg aattggaacg tgaagaatta tggaaaaaat
     3481 tagaggagct aaagctaaag aaagctctag aaaaacagaa tagtgcttac aacatgcaca
     3541 gtattctcag caatacaagt gccgaagttt aaacgaattc gggctcggta cccggggatc
     3601 ctctagagtc gacctgcagg catgcaagct gatttggctg ctgccaccgc tgagcaaggc
     3661 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3721 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3781 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3841 gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcttaa
     3901 ttaagaattc taatataact tcgtatagca tacattatac gaagttatgg atccactgtt
     3961 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     4021 acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4081 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4141 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4201 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4261 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4321 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4381 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4441 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4501 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4561 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4621 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4681 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4741 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4801 cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag cgacgcccaa
     4861 ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4921 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5401 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa
     5581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5641 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaatc
     5701 gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat tgcggccatc
     5761 gtgatggcta gagtttagcc ctcccacaca taaccagagg gcagcaattc acgaatccca
     5821 actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg atgcagatcg
     5881 agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc atttccgatc
     5941 gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag caccacgagt
     6001 tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat gcggccgtcg
     6061 ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct gttgattgta
     6121 gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc catggtggcg
     6181 ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac
     6241 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt
     6301 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
     6361 gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     6421 gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc
     6481 atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac
     6541 agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga ggttatttca
     6601 ggccatggtg ctgcgcaaga cgcgttaatt aagacgt
//
