LOCUS       sequence-475192-        6966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express bTrCP-HaloTag 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 6966)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6966)
  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            "Unpublished"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: bTrCP-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..6966
                     /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"
     CDS             2901..2921
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2937..3821
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(3909..3927)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3964..4085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3975..3994)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4029..4048
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4330..4687
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4330..4521
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4353..4373)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4538..4673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4702..4749
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4781..5575
                     /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(4835..4854)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5445..5464
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5639..5687
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5926..6514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6006..6025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6630..6913
                     /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 ggctagcgat cgccatggac ccggccgagg
     1081 cggtgctgca agagaaggca ctcaagttta tgtgctctat gcccaggtct ctgtggctgg
     1141 gctgctccag cctggcggac agcatgcctt cgctgcgatg cctgtataac ccagggactg
     1201 gcgcactcac agctttccag aattcctcag agagagaaga ctgtaataat ggcgaacccc
     1261 ctaggaagat aataccagag aagaattcac ttagacagac atacaacagc tgtgccagac
     1321 tctgcttaaa ccaagaaaca gtatgtttag caagcactgc tatgaagact gagaattgtg
     1381 tggccaaaac aaaacttgcc aatggcactt ccagtatgat tgtgcccaag caacggaaac
     1441 tctcagcaag ctatgaaaag gaaaaggaac tgtgtgtcaa atactttgag cagtggtcag
     1501 agtcagatca agtggaattt gtggaacatc ttatatccca aatgtgtcat taccaacatg
     1561 ggcacataaa ctcgtatctt aaacctatgt tgcagagaga tttcataact gctctgccag
     1621 ctcggggatt ggatcatatt gctgagaaca ttctgtcata cctggatgcc aaatcactat
     1681 gtgctgctga acttgtgtgc aaggaatggt accgagtgac ctctgatggc atgctgtgga
     1741 agaagcttat cgagagaatg gtcaggacag attctctgtg gagaggcctg gcagaacgaa
     1801 gaggatgggg acagtattta ttcaaaaaca aacctcctga cgggaatgct cctcccaact
     1861 ctttttatag agcactttat cctaaaatta tacaagacat tgagacaata gaatctaatt
     1921 ggagatgtgg aagacatagt ttacagagaa ttcactgccg aagtgaaaca agcaaaggag
     1981 tttactgttt acagtatgat gatcagaaaa tagtaagcgg ccttcgagac aacacaatca
     2041 agatctggga taaaaacaca ttggaatgca agcgaattct cacaggccat acaggttcag
     2101 tcctctgtct ccagtatgat gagagagtga tcataacagg atcatcggat tccacggtca
     2161 gagtgtggga tgtaaataca ggtgaaatgc taaacacgtt gattcaccat tgtgaagcag
     2221 ttctgcactt gcgtttcaat aatggcatga tggtgacctg ctccaaagat cgttccattg
     2281 ctgtatggga tatggcctcc ccaactgaca ttaccctccg gagggtgctg gtcggacacc
     2341 gagctgctgt caatgttgta gactttgatg acaagtacat tgtttctgca tctggggata
     2401 gaactataaa ggtatggaac acaagtactt gtgaatttgt aaggacctta aatggacaca
     2461 aacgaggcat tgcctgtttg cagtacaggg acaggctggt agtgagtggc tcatctgaca
     2521 acactatcag attatgggac atagaatgtg gtgcatgttt acgagtgtta gaaggccatg
     2581 aggaattggt gcgttgtatt cgatttgata acaagaggat agtcagtggg gcctatgatg
     2641 gaaaaattaa agtgtgggat cttgtggctg ctttggaccc ccgtgctcct gcagggacac
     2701 tctgtctacg gacccttgtg gagcattccg gaagagtttt tcgactacag tttgatgaat
     2761 tccagattgt cagtagttca catgatgaca caatcctcat ctgggacttc ctaaatgatc
     2821 cagctgccca agctgaaccc ccccgttccc cttctcgaac atacacctac atctccagag
     2881 tttctctcga gccaaccact gaggatctgt actttcagag cgataacgat ggatccgaaa
     2941 tcggtactgg ctttccattc gacccccatt atgtggaagt cctgggcgag cgcatgcact
     3001 acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt cctgcacggt aacccgacct
     3061 cctcctacgt gtggcgcaac atcatcccgc atgttgcacc gacccatcgc tgcattgctc
     3121 cagacctgat cggtatgggc aaatccgaca aaccagacct gggttatttc ttcgacgacc
     3181 acgtccgctt catggatgcc ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca
     3241 ttcacgactg gggctccgct ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca
     3301 aaggtattgc atttatggag ttcatccgcc ctatcccgac ctgggacgaa tggccagaat
     3361 ttgcccgcga gaccttccag gccttccgca ccaccgacgt cggccgcaag ctgatcatcg
     3421 atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag
     3481 tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc
     3541 gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag
     3601 aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg ggcaccccag
     3661 gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct aactgcaagg
     3721 ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac ctgatcggca
     3781 gcgagatcgc gcgctggctg tctactctgg agatttccgg ttaatagaat tctagagtcg
     3841 acctgcaggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     3901 tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt cgagcagaca
     3961 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4021 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4081 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     4141 tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa caaaatatta
     4201 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4261 gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt
     4321 aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4381 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4441 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4501 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4561 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4621 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4681 ttgcaaaaag cttaattaac tgttgacaat taatcatcgg catagtatat cggcatagta
     4741 taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt
     4801 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4861 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4921 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4981 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5041 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5101 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5161 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5221 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5281 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5341 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5401 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5461 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5521 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5581 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa
     5641 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat
     5701 aaggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     5761 ccggggtcag caccgtttct gcggactggc tttctacccg gtatcagctc actcaaaggc
     5821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6361 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6481 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     6541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6601 aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc
     6661 cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat
     6721 gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt
     6781 gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg
     6841 aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc
     6901 tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg
     6961 agatct
//
