LOCUS       sequence-479885-        6358 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HaloTag-p53 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 6358)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6358)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HaloTag-p53 Vector
FEATURES             Location/Qualifiers
     source          1..6358
                     /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"
     CDS             2109..3293
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSDV"
     polyA_signal    3400..3521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3411..3430)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3636..3669)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3679..3726
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3758..4552
                     /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(3812..3831)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4422..4441
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4710..5298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4790..4809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5424..5472
                     /note="synthetic polyadenylation signal"
     CDS             complement(5496..5894)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5891..5900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5925..6282)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5939..6074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6091..6282
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6239..6259
                     /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 ggaggagccg cagtcagatc ctagcgtcga gccccctctg agtcaggaaa
     2161 cattttcaga cctatggaaa ctacttcctg aaaacaacgt tctgtccccc ttgccgtccc
     2221 aagcaatgga tgatttgatg ctgtccccgg acgatattga acaatggttc actgaagacc
     2281 caggtccaga tgaagctccc agaatgccag aggctgctcc ccccgtggcc cctgcaccag
     2341 cagctcctac accggcggcc cctgcaccag ccccctcctg gcccctgtca tcttctgtcc
     2401 cttcccagaa aacctaccag ggcagctacg gtttccgtct gggcttcttg cattctggga
     2461 cagccaagtc tgtgacttgc acgtactccc ctgccctcaa caagatgttt tgccaactgg
     2521 ccaagacctg ccctgtgcag ctgtgggttg attccacacc cccgcccggc acccgcgtcc
     2581 gcgccatggc catctacaag cagtcacagc acatgacgga ggttgtgagg cgctgccccc
     2641 accatgagcg ctgctcagat agcgatggtc tggcccctcc tcagcatctt atccgagtgg
     2701 aaggaaattt gcgtgtggag tatttggatg acagaaacac ttttcgacat agtgtggtgg
     2761 tgccctatga gccgcctgag gttggctctg actgtaccac catccactac aactacatgt
     2821 gtaacagttc ctgcatgggc ggcatgaacc ggaggcccat cctcaccatc atcacactgg
     2881 aagactccag tggtaatcta ctgggacgga acagctttga ggtgcgtgtt tgtgcctgtc
     2941 ctgggagaga ccggcgcaca gaggaagaga atctccgcaa gaaaggggag cctcaccacg
     3001 agctgccccc agggagcact aagcgagcac tgcccaacaa caccagctcc tctccccagc
     3061 caaagaagaa accactggat ggagaatatt tcacccttca gatccgtggg cgtgagcgct
     3121 tcgagatgtt ccgagagctg aatgaggcct tggaactcaa ggatgcccag gctgggaagg
     3181 agccaggggg gagcagggct cactccagcc acctgaagtc caaaaagggt cagtctacct
     3241 cccgccataa aaaactcatg ttcaagacag aagggcctga ctcagacgtt taaacgaatt
     3301 cgggctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc tgatttggct
     3361 gctgccaccg ctgagcaagg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     3421 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3481 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3541 attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct
     3601 acaaatgtgg taaaatctta attaagaatt ctaatataac ttcgtatagc atacattata
     3661 cgaagttatg gatccactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3721 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     3781 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3841 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3901 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3961 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4021 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4081 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4141 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4201 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4261 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4321 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4381 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4441 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4501 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gattcgaaat
     4561 gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc
     4621 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5281 agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5401 atccttttat agtccggaat cgtcacacaa aaaaccaaca cacagatgta atgaaaataa
     5461 agatatttta ttgcggccat cgtgatggct agagtttagc cctcccacac ataaccagag
     5521 ggcagcaatt cacgaatccc aactgccgtc ggctgtccat cactgtcctt cactatggct
     5581 ttgatcccag gatgcagatc gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg
     5641 cccctgttct catttccgat cgcgacgata caagtcaggt tgccagctgc cgcagcagca
     5701 gcagtgccca gcaccacgag ttctgcacaa ggtcccccag taaaatgata tacattgaca
     5761 ccagtgaaga tgcggccgtc gctagagaga gctgcgctgg cgacgctgta gtcttcagag
     5821 atggggatgc tgttgattgt agccgttgct ctttcaatga gggtggattc ttcttgagac
     5881 aaaggcttgg ccatggtggc gctagtgtca gaagaatcga gctttttgca aaagcctagg
     5941 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     6001 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     6061 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     6121 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     6181 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     6241 cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc taaccaagtt
     6301 cctctttcag aggttatttc aggccatggt gctgcgcaag acgcgttaat taagacgt
//
