LOCUS       sequence-479220-        6307 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HaloTag-PSMD3 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 6307)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6307)
  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            "Unpublished"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: NanoBRET Assay Vector, HaloTag-PSMD3
FEATURES             Location/Qualifiers
     source          1..6307
                     /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(3721..3739)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3776..3897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3787..3806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3841..3860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4144..4248
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4249..4317,4318..5109)
                     /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(4467..4486)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5267..5855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5347..5366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5971..6254
                     /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 tcgccatgaa gcaggagggc tcggcgcggc gccgcggcgc
     2041 ggacaaggcg aaaccgccgc ccggcggagg agaacaagaa cccccaccgc cgccggcccc
     2101 ccaggatgtg gagatgaaag aggaggcagc gacgggtggc gggtcaacgg gggaggcaga
     2161 cggcaagacg gcggcggcag cggctgagca ctcccagcga gagctggaca cagtcacctt
     2221 ggaggacatc aaggagcacg tgaaacagct agagaaagcg gtttcaggca aggagccgag
     2281 attcgtgctg cgggccctgc ggatgctgcc ttccacatca cgccgcctca accactatgt
     2341 tctgtataag gctgtgcagg gcttcttcac ttcaaataat gccactcgag actttttgct
     2401 ccccttcctg gaagagccca tggacacaga ggctgattta cagttccgtc cccgcacggg
     2461 aaaagctgcg tcgacacccc tcctgcctga agtggaagcc tatctccaac tcctcgtggt
     2521 catcttcatg atgaacagca agcgctacaa agaggcacag aagatctctg atgatctgat
     2581 gcagaagatc agtactcaga accgccgggc cctagacctt gtagccgcaa agtgttacta
     2641 ttatcacgcc cgggtctatg agttcctgga caagctggat gtggtgcgca gcttcttgca
     2701 tgctcggctc cggacagcta cgcttcggca tgacgcagac gggcaggcca ccctgttgaa
     2761 cctcctgctg cggaattacc tacactacag cttgtacgac caggctgaga agctggtgtc
     2821 caagtctgtg ttcccagagc aggccaacaa caatgagtgg gccaggtacc tctactacac
     2881 agggcgaatc aaagccatcc agctggagta ctcagaggcc cggagaacga tgaccaacgc
     2941 ccttcgcaag gcccctcagc acacagctgt cggcttcaaa cagacggtgc acaagcttct
     3001 catcgtggtg gagctgttgc tgggggagat ccctgaccgg ctgcagttcc gccagccctc
     3061 cctcaagcgc tcactcatgc cctatttcct tctgactcaa gctgtcagga caggaaacct
     3121 agccaagttc aaccaggtcc tggatcagtt tggggagaag tttcaagcag atgggaccta
     3181 caccctaatt atccggctgc ggcacaacgt gattaagaca ggtgtacgca tgatcagcct
     3241 ctcctattcc cgaatctcct tggctgacat cgcccagaag ctgcagttgg atagccccga
     3301 agatgcagag ttcattgttg ccaaggccat ccgggatggt gtcattgagg ccagcatcaa
     3361 ccacgagaag ggctatgtcc aatccaagga gatgattgac atctattcca cccgagagcc
     3421 ccagctagcc ttccaccagc gcatctcctt ctgcctagat atccacaaca tgtctgtcaa
     3481 ggccatgagg tttcctccca aatcgtacaa caaggacttg gagtctgcag aggaacggcg
     3541 tgagcgagaa cagcaggact tggagtttgc caaggagatg gcagaagatg atgatgacag
     3601 cttccctgtt taaacgaatt cgggctcggt acccggggat cctctagagt cgacctgcag
     3661 gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     3721 ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga
     3781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3901 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta
     3961 agcaagtaaa acctctacaa atgtggtaaa atcgaattct aatggatcct ctttgcgctt
     4021 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     4081 tgcggactgg ctttctacgt aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4141 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4201 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4261 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4321 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4381 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4441 ccaatgatga gcactttcaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4501 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4561 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4621 ataaccatga gtgataacac tgcggccaac ttacttctga caactatcgg aggaccgaag
     4681 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4741 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4801 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4861 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4921 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4981 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5041 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5101 cattggtaat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     5161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5701 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5821 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5941 aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc
     6001 ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa
     6061 tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat
     6121 tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct
     6181 gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     6241 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     6301 gagatct
//
