LOCUS       sequence-479216-        6520 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HaloTag-bTrCP 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 6520)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6520)
  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            "Unpublished"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-bTrCP Vector
FEATURES             Location/Qualifiers
     source          1..6520
                     /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(3934..3952)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3989..4110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4000..4019)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4054..4073
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4357..4461
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4462..4530,4531..5322)
                     /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(4680..4699)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5480..6068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5560..5579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6184..6467
                     /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 tcgccatgga cccggccgag gcggtgctgc aagagaaggc
     2041 actcaagttt atgtgctcta tgcccaggtc tctgtggctg ggctgctcca gcctggcgga
     2101 cagcatgcct tcgctgcgat gcctgtataa cccagggact ggcgcactca cagctttcca
     2161 gaattcctca gagagagaag actgtaataa tggcgaaccc cctaggaaga taataccaga
     2221 gaagaattca cttagacaga catacaacag ctgtgccaga ctctgcttaa accaagaaac
     2281 agtatgttta gcaagcactg ctatgaagac tgagaattgt gtggccaaaa caaaacttgc
     2341 caatggcact tccagtatga ttgtgcccaa gcaacggaaa ctctcagcaa gctatgaaaa
     2401 ggaaaaggaa ctgtgtgtca aatactttga gcagtggtca gagtcagatc aagtggaatt
     2461 tgtggaacat cttatatccc aaatgtgtca ttaccaacat gggcacataa actcgtatct
     2521 taaacctatg ttgcagagag atttcataac tgctctgcca gctcggggat tggatcatat
     2581 tgctgagaac attctgtcat acctggatgc caaatcacta tgtgctgctg aacttgtgtg
     2641 caaggaatgg taccgagtga cctctgatgg catgctgtgg aagaagctta tcgagagaat
     2701 ggtcaggaca gattctctgt ggagaggcct ggcagaacga agaggatggg gacagtattt
     2761 attcaaaaac aaacctcctg acgggaatgc tcctcccaac tctttttata gagcacttta
     2821 tcctaaaatt atacaagaca ttgagacaat agaatctaat tggagatgtg gaagacatag
     2881 tttacagaga attcactgcc gaagtgaaac aagcaaagga gtttactgtt tacagtatga
     2941 tgatcagaaa atagtaagcg gccttcgaga caacacaatc aagatctggg ataaaaacac
     3001 attggaatgc aagcgaattc tcacaggcca tacaggttca gtcctctgtc tccagtatga
     3061 tgagagagtg atcataacag gatcatcgga ttccacggtc agagtgtggg atgtaaatac
     3121 aggtgaaatg ctaaacacgt tgattcacca ttgtgaagca gttctgcact tgcgtttcaa
     3181 taatggcatg atggtgacct gctccaaaga tcgttccatt gctgtatggg atatggcctc
     3241 cccaactgac attaccctcc ggagggtgct ggtcggacac cgagctgctg tcaatgttgt
     3301 agactttgat gacaagtaca ttgtttctgc atctggggat agaactataa aggtatggaa
     3361 cacaagtact tgtgaatttg taaggacctt aaatggacac aaacgaggca ttgcctgttt
     3421 gcagtacagg gacaggctgg tagtgagtgg ctcatctgac aacactatca gattatggga
     3481 catagaatgt ggtgcatgtt tacgagtgtt agaaggccat gaggaattgg tgcgttgtat
     3541 tcgatttgat aacaagagga tagtcagtgg ggcctatgat ggaaaaatta aagtgtggga
     3601 tcttgtggct gctttggacc cccgtgctcc tgcagggaca ctctgtctac ggacccttgt
     3661 ggagcattcc ggaagagttt ttcgactaca gtttgatgaa ttccagattg tcagtagttc
     3721 acatgatgac acaatcctca tctgggactt cctaaatgat ccagctgccc aagctgaacc
     3781 cccccgttcc ccttctcgaa catacaccta catctccaga gtttaaacga attcgggctc
     3841 ggtacccggg gatcctctag agtcgacctg caggcatgca agctgatccg gctgctaaca
     3901 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     3961 ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     4021 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4081 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4141 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     4201 aaaatcgaat tctaatggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag
     4261 tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtaatggtt
     4321 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4381 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4441 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4501 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4561 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4621 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt caaagttctg
     4681 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4741 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4801 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4861 aacttacttc tgacaactat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4921 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4981 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5041 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5101 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5161 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5221 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5281 cagatcgctg agataggtgc ctcactgatt aagcattggt aattcgaaat gaccgaccaa
     5341 gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5401 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5641 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5881 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6061 gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6121 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttat
     6181 agtccggaaa tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa
     6241 aaatggcagc ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag
     6301 gtattcatac ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg
     6361 ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt
     6421 aagggattat ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt
     6481 actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
