LOCUS       sequence-481754-        6196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag-MDM2 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 6196)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6196)
  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            "Unpublished"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: HaloTag-MDM2 Vector
FEATURES             Location/Qualifiers
     source          1..6196
                     /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"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /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             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(3610..3628)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3665..3786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3676..3695)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3730..3749
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4033..4137
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4138..4206,4207..4998)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4356..4375)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5156..5744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5236..5255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5860..6143
                     /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 ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgccatggt gaggagcagg caaatgtgca ataccaacat
     2041 gtctgtacct actgatggtg ctgtaaccac ctcacagatt ccagcttcgg aacaagagac
     2101 cctggttaga ccaaagccat tgcttttgaa gttattaaag tctgttggtg cacaaaaaga
     2161 cacttatact atgaaagagg ttctttttta tcttggccag tatattatga ctaaacgatt
     2221 atatgatgag aagcaacaac atattgtata ttgttcaaat gatcttctag gagatttgtt
     2281 tggcgtgcca agcttctctg tgaaagagca caggaaaata tataccatga tctacaggaa
     2341 cttggtagta gtcaatcagc aggaatcatc ggactcaggt acatctgtga gtgagaacag
     2401 gtgtcacctt gaaggtggga gtgatcaaaa ggaccttgta caagagcttc aggaagagaa
     2461 accttcatct tcacatttgg tttctagacc atctacctca tctagaagga gagcaattag
     2521 tgagacagaa gaaaattcag atgaattatc tggtgaacga caaagaaaac gccacaaatc
     2581 tgatagtatt tccctttcct ttgatgaaag cctggctctg tgtgtaataa gggagatatg
     2641 ttgtgaaaga agcagtagca gtgaatctac agggacgcca tcgaatccgg atcttgatgc
     2701 tggtgtaagt gaacattcag gtgattggtt ggatcaggat tcagtttcag atcagtttag
     2761 tgtagaattt gaagttgaat ctctcgactc agaagattat agccttagtg aagaaggaca
     2821 agaactctca gatgaagatg atgaggtata tcaagttact gtgtatcagg caggggagag
     2881 tgatacagat tcatttgaag aagatcctga aatttcctta gctgactatt ggaaatgcac
     2941 ttcatgcaat gaaatgaatc ccccccttcc atcacattgc aacagatgtt gggcccttcg
     3001 tgagaattgg cttcctgaag ataaagggaa agataaaggg gaaatctctg agaaagccaa
     3061 actggaaaac tcaacacaag ctgaagaggg ctttgatgtt cctgattgta aaaaaactat
     3121 agtgaatgat tccagagagt catgtgttga ggaaaatgat gataaaatta cacaagcttc
     3181 acaatcacaa gaaagtgaag actattctca gccatcaact tctagtagca ttatttatag
     3241 cagccaagaa gatgtgaaag agtttgaaag ggaagaaacc caagacaaag aagagagtgt
     3301 ggaatctagt ttgcccctta atgccattga accttgtgtg atttgtcaag gtcgacctaa
     3361 aaatggttgc attgtccatg gcaaaacagg acatcttatg gcctgcttta catgtgcaaa
     3421 gaagctaaag aaaaggaata agccctgccc agtatgtaga caaccaattc aaatgattgt
     3481 gctaacttat ttccccgttt aaacgaattc gggctcggta cccggggatc ctctagagtc
     3541 gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     3601 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     3661 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3721 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3781 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     3841 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaattcta atggatcctc
     3901 tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag
     3961 caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt
     4021 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4081 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4141 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4201 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4261 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4321 gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt
     4381 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4441 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4501 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aactatcgga
     4561 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4621 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4681 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4741 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4801 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4861 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4921 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4981 ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     5041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5101 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5341 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5521 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5581 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5641 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5701 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     5761 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5821 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg
     5881 ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag
     5941 tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc
     6001 aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt
     6061 gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta
     6121 ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg
     6181 tagaggatcg agatct
//
