LOCUS       sequence-479778-        6951 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express DCAF15-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 6951)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6951)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: DCAF15-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..6951
                     /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             2886..2906
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2922..3806
                     /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(3894..3912)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3949..4070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3960..3979)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4014..4033
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4315..4672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4315..4506
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4338..4358)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4523..4658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4687..4734
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4766..5560
                     /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(4820..4839)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5430..5449
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5624..5672
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5911..6499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5991..6010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6615..6898
                     /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 cgccatggct ccatcaagca
     1081 aaagcgaaag gaattccggc gcgggctccg gtggaggagg tcccggcgga gctggaggaa
     1141 agcgagccgc tggtagaagg cgagaacacg tacttaaaca gcttgaacga gttaagattt
     1201 ccgggcaact ctcaccgcgg ttgtttagga aactgccccc tagagtatgt gtgtccttga
     1261 agaacattgt ggacgaagat ttcctttacg ctgggcacat ctttctcggc ttttcaaaat
     1321 gtgggagata cgtgctttca tacaccagca gttcaggcga tgacgacttc tcattctata
     1381 tctaccattt gtattggtgg gagttcaatg tacattctaa gcttaaactt gtgcgacagg
     1441 tgaggctgtt tcaggatgag gagatatatt ccgaccttta tttgacagtt tgcgagtggc
     1501 cgtctgatgc ttccaaagtg atagtttttg gatttaatac tcggagcgcc aacggaatgc
     1561 ttatgaacat gatgatgatg tctgatgaaa atcatcgaga catttatgtc tcaacagtcg
     1621 ctgttccacc gcctggacgc tgcgctgcat gccaggacgc gagcagggcc caccccggtg
     1681 atccaaacgc ccaatgtctt cgccacgggt ttatgttgca tacgaagtac caggtcgtgt
     1741 atcccttccc cactttccaa cccgcgttcc aactgaaaaa agatcaagtc gttctgttga
     1801 acacgtcata tagtttggtt gcttgtgcgg tctctgttca tagtgcggga gacaggtctt
     1861 tctgtcaaat cctgtatgat cactccacct gtcctctcgc gccagcgtcc cctcccgagc
     1921 cacagagccc cgaactgcct ccagcacttc cttccttctg tcctgaggct gctccagccc
     1981 gctcaagcgg tagcccggag ccgtcccccg ctatcgcgaa ggcaaaagaa tttgttgcgg
     2041 acatcttcag gcgggctaag gaagcaaagg ggggtgttcc cgaggaggcc aggcccgcgc
     2101 tgtgtcctgg gccttctggg agtcgatgcc gcgcgcatag tgaaccactg gcactctgtg
     2161 gggagaccgc cccacgcgac tcacccccag cgtctgaggc accggcgtca gaaccgggct
     2221 atgtcaatta cactaaactt tattacgttc ttgaaagtgg agaggggaca gagcccgaag
     2281 acgaacttga ggatgacaag atcagcctgc cgtttgtagt gacggacttg cgcgggcgaa
     2341 atttgcggcc tatgagggaa cgaaccgccg tccaagggca gtaccttact gttgaacagc
     2401 tcactttgga ctttgagtat gtaataaacg aggtaattag acatgacgct acatggggcc
     2461 atcagttctg ctcattcagt gattatgata ttgtaatact ggaagtgtgc cctgaaacaa
     2521 atcaagtcct tataaatatc gggctgttgt tgctcgcgtt tccgagtcct actgaagaag
     2581 ggcagcttcg accgaagacc taccatacgt ctcttaaagt cgcatgggac ctcaacaccg
     2641 gcatcttcga gaccgtgagc gtgggcgatt tgacagaagt gaaaggacaa acaagcggtt
     2701 ctgtttggtc ctcctatcgg aaaagttgcg tagacatggt catgaagtgg ttggttccag
     2761 agagtagcgg tagatatgtg aatcggatga ccaatgaggc tttgcataag ggctgctctt
     2821 tgaaagtgct ggcagacagc gagcgatata cttggatagt cctggtttct ctcgagccaa
     2881 ccactgagga tctgtacttt cagagcgata acgatggatc cgaaatcggt actggctttc
     2941 cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc
     3001 cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc
     3061 gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta
     3121 tgggcaaatc cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg
     3181 atgccttcat cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct
     3241 ccgctctggg tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta
     3301 tggagttcat ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct
     3361 tccaggcctt ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta
     3421 tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt
     3481 accgcgagcc gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc
     3541 tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc
     3601 tgcaccagtc ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac
     3661 cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc
     3721 cgggtctgaa tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct
     3781 ggctgtctac tctggagatt tccggttaat agaattctag agtcgacctg caggcatgca
     3841 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     3901 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     3961 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4021 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4081 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     4141 aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc
     4201 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     4261 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     4321 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4381 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4441 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4501 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4561 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4621 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa
     4681 ttaactgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     4741 aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc
     4801 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4861 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4921 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4981 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5041 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5101 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5161 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5221 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5281 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5341 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5401 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5461 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5521 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5581 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     5641 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tcctctttgc
     5701 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
     5761 tttctgcgga ctggctttct acccggtatc agctcactca aaggcggtaa tacggttatc
     5821 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5881 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5941 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6001 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6061 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6121 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6181 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6241 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6301 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6361 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6421 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6481 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6541 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6601 gatcctttta tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt
     6661 gaaaccatga aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc
     6721 tggttgcata ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg
     6781 gtggtttgtt gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg
     6841 tgcgtacaat taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga
     6901 taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc t
//
