LOCUS       sequence-481983-        6246 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express DCAF1(del)-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 6246)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6246)
  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            "Unpublished"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: DCAF1(del)-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..6246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3..84
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        317..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          960..1092
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1075..1094
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1136..1155
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1136..1154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2281..2301
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2317..3201
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    3288..3409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3299..3318)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3524..3557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3567..3614
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3646..4440
                     /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(3700..3719)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4310..4329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4598..5186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4678..4697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5312..5360
                     /note="synthetic polyadenylation signal"
     CDS             complement(5384..5782)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5779..5788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5813..6170)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5827..5962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5979..6170
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6127..6147
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 cggcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
       61 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      121 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
      181 gattatgatc cgcggccgca atacgtcgac gttatcagct gacttcgtac gagagcctag
      241 gattatggcg cgccactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      841 agatcactag aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      901 cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg
      961 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1021 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     1081 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtattaata
     1141 cgactcacta tagggctagc gatcgccatg gagccaaaac agaggcggca agcgccaata
     1201 aactttacgt caaggctaaa ccgcagggca tcatttccaa agtatggagg ggtggatggc
     1261 ggatgctttg ataggcacct tatctttagc agattccgtc ctatttcagt gttccgggaa
     1321 gccaatgaag atgagagtgg cttcacctgc tgtgcattct cagcacggga gcggttcctg
     1381 atgcttggca cctgcacagg gcagctgaag ctctataatg tgtttagtgg acaggaggag
     1441 gccagctata actgtcacaa ctcagccatc acacatcttg aaccttccag ggatgggtcc
     1501 ttgctgctga catctgctac ttggagccag cctttgtctg cactttgggg aatgaagtca
     1561 gtatttgata tgaagcattc cttcacagaa gatcactatg ttgagttcag taagcactcc
     1621 caggatcggg tcatcggcac aaaaggagac attgcccaca tttatgatat tcagactggc
     1681 aacaagctgt tgactctgtt taacccagat cttgccaaca actacaagag gaactgtgcc
     1741 acctttaatc ctacagatga tcttgtctta aatgatggcg tcctctggga tgtccgctct
     1801 gcacaggcca tccacaagtt tgacaagttc aatatgaaca tcagtggtgt tttccatcca
     1861 aatggactgg aggtgatcat taatactgag atttgggacc ttcgaacttt tcatcttttg
     1921 catactgttc ccgctctgga tcagtgtcgc gtggtgttca atcacacggg aacagtgatg
     1981 tatggagcta tgttgcaggc agatgatgaa gatgacttaa tggaagagag gatgaaaagc
     2041 ccctttgggt catccttccg aacatttaat gcaactgact acaaacctat agcaaccatt
     2101 gatgtgaaac ggaacatctt tgacctgtgt acagacacca aagactgcta tcttgctgtc
     2161 attgagaatc aaggcagcat ggatgccctg aacatggaca cagtatgcag gctgtatgaa
     2221 gtgggcaggc agcgtctggc agaggatgag gatgaagagg tttctctcga gccaaccact
     2281 gaggatctgt actttcagag cgataacgat ggatccgaaa tcggtactgg ctttccattc
     2341 gacccccatt atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc
     2401 gatggcaccc ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac
     2461 atcatcccgc atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc
     2521 aaatccgaca aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc
     2581 ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct
     2641 ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag
     2701 ttcatccgcc ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag
     2761 gccttccgca ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag
     2821 ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc
     2881 gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca
     2941 atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac
     3001 cagtcccctg tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc
     3061 gaagccgctc gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt
     3121 ctgaatctgc tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg
     3181 tctactctgg agatttccgg ttaatagaat tctagagtcg acctgcaggc atgcaagctg
     3241 atttggctgc tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga
     3301 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3361 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3421 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     3481 aaacctctac aaatgtggta aaatcttaat taagaattct aatataactt cgtatagcat
     3541 acattatacg aagttatgga tccactgttg acaattaatc atcggcatag tatatcggca
     3601 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     3661 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3721 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3781 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3841 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3901 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3961 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4021 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4081 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4141 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4201 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4261 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4321 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4381 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4441 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     4501 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4561 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4621 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4681 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4741 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4801 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4861 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4921 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4981 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5041 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5101 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5161 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     5221 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5281 tcacctagat ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat
     5341 gaaaataaag atattttatt gcggccatcg tgatggctag agtttagccc tcccacacat
     5401 aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca
     5461 ctatggcttt gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc
     5521 tcaagatgcc cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg
     5581 cagcagcagc agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata
     5641 cattgacacc agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt
     5701 cttcagagat ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt
     5761 cttgagacaa aggcttggcc atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa
     5821 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     5881 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     5941 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     6001 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     6061 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     6121 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta
     6181 accaagttcc tctttcagag gttatttcag gccatggtgc tgcgcaagac gcgttaatta
     6241 agacgt
//
