LOCUS       sequence-479218-        7973 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HaloTag-DCAF15 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 7973)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7973)
  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            "Unpublished"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-DCAF15 Vector
FEATURES             Location/Qualifiers
     source          1..7973
                     /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(3919..3937)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3974..4095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3985..4004)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4039..4058
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4340..4697
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4340..4531
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4363..4383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4548..4683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4748..5542
                     /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(4802..4821)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5412..5431
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5606..5654
                     /note="synthetic polyadenylation signal"
     promoter        5810..5914
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5915..5983,5984..6775)
                     /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(6133..6152)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6933..7521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7013..7032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7637..7920
                     /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 tcgccatggc tccatcaagc aaaagcgaaa ggaattccgg
     2041 cgcgggctcc ggtggaggag gtcccggcgg agctggagga aagcgagccg ctggtagaag
     2101 gcgagaacac gtacttaaac agcttgaacg agttaagatt tccgggcaac tctcaccgcg
     2161 gttgtttagg aaactgcccc ctagagtatg tgtgtccttg aagaacattg tggacgaaga
     2221 tttcctttac gctgggcaca tctttctcgg cttttcaaaa tgtgggagat acgtgctttc
     2281 atacaccagc agttcaggcg atgacgactt ctcattctat atctaccatt tgtattggtg
     2341 ggagttcaat gtacattcta agcttaaact tgtgcgacag gtgaggctgt ttcaggatga
     2401 ggagatatat tccgaccttt atttgacagt ttgcgagtgg ccgtctgatg cttccaaagt
     2461 gatagttttt ggatttaata ctcggagcgc caacggaatg cttatgaaca tgatgatgat
     2521 gtctgatgaa aatcatcgag acatttatgt ctcaacagtc gctgttccac cgcctggacg
     2581 ctgcgctgca tgccaggacg cgagcagggc ccaccccggt gatccaaacg cccaatgtct
     2641 tcgccacggg tttatgttgc atacgaagta ccaggtcgtg tatcccttcc ccactttcca
     2701 acccgcgttc caactgaaaa aagatcaagt cgttctgttg aacacgtcat atagtttggt
     2761 tgcttgtgcg gtctctgttc atagtgcggg agacaggtct ttctgtcaaa tcctgtatga
     2821 tcactccacc tgtcctctcg cgccagcgtc ccctcccgag ccacagagcc ccgaactgcc
     2881 tccagcactt ccttccttct gtcctgaggc tgctccagcc cgctcaagcg gtagcccgga
     2941 gccgtccccc gctatcgcga aggcaaaaga atttgttgcg gacatcttca ggcgggctaa
     3001 ggaagcaaag gggggtgttc ccgaggaggc caggcccgcg ctgtgtcctg ggccttctgg
     3061 gagtcgatgc cgcgcgcata gtgaaccact ggcactctgt ggggagaccg ccccacgcga
     3121 ctcaccccca gcgtctgagg caccggcgtc agaaccgggc tatgtcaatt acactaaact
     3181 ttattacgtt cttgaaagtg gagaggggac agagcccgaa gacgaacttg aggatgacaa
     3241 gatcagcctg ccgtttgtag tgacggactt gcgcgggcga aatttgcggc ctatgaggga
     3301 acgaaccgcc gtccaagggc agtaccttac tgttgaacag ctcactttgg actttgagta
     3361 tgtaataaac gaggtaatta gacatgacgc tacatggggc catcagttct gctcattcag
     3421 tgattatgat attgtaatac tggaagtgtg ccctgaaaca aatcaagtcc ttataaatat
     3481 cgggctgttg ttgctcgcgt ttccgagtcc tactgaagaa gggcagcttc gaccgaagac
     3541 ctaccatacg tctcttaaag tcgcatggga cctcaacacc ggcatcttcg agaccgtgag
     3601 cgtgggcgat ttgacagaag tgaaaggaca aacaagcggt tctgtttggt cctcctatcg
     3661 gaaaagttgc gtagacatgg tcatgaagtg gttggttcca gagagtagcg gtagatatgt
     3721 gaatcggatg accaatgagg ctttgcataa gggctgctct ttgaaagtgc tggcagacag
     3781 cgagcgatat acttggatag tcctggttta aacgaattcg ggctcggtac ccggggatcc
     3841 tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag
     3901 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt
     3961 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4021 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4081 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4141 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa
     4201 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     4261 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
     4321 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4501 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4561 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4621 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4681 gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac ttaaggctag
     4741 agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4801 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4861 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4921 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4981 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5041 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5101 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5161 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5221 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5281 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5341 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5401 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5461 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5521 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5581 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5641 tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt
     5701 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     5761 ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5821 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5881 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5941 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6001 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6061 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6121 tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6181 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6241 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6301 taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc taaccgcttt
     6361 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6421 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6481 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6541 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6601 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6661 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6721 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaattcga
     6781 aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta
     6841 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6901 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6961 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7021 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7081 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7141 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7201 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7261 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7321 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7381 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7441 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7501 cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7561 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7621 tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg
     7681 gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc
     7741 tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc
     7801 gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc
     7861 ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga
     7921 gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
