LOCUS       sequence-473352-        6511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag(R)-hVPS4B 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 6511)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6511)
  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            "Unpublished"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: HaloTag-hVPS4B Vector
FEATURES             Location/Qualifiers
     source          1..6511
                     /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"
     regulatory      1164..1173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1170..2060
                     /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             2073..2093
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2932..2943
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    3553..3674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3564..3583)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3789..3822)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3832..3879
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3911..4705
                     /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(3965..3984)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4575..4594
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4863..5451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4943..4962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5577..5625
                     /note="synthetic polyadenylation signal"
     CDS             complement(5649..6047)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6044..6053
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6078..6435)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6092..6227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6244..6435
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6392..6412
                     /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 aaagccacca tggcagaaat cggtactggc tttccattcg
     1201 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     1261 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     1321 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     1381 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     1441 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     1501 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     1561 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     1621 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     1681 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     1741 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     1801 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     1861 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     1921 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     1981 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     2041 cgacgctcga gatttccggc gagccaacca ctgaggatct gtactttcag agcgataacg
     2101 cgatcgccat gtcatccact tcgcccaacc tccagaaagc gatagatctg gctagcaaag
     2161 cagcgcaaga agacaaggct gggaactacg aagaagccct tcagctctat cagcatgctg
     2221 tgcagtattt tcttcatgtc gttaaatatg aagcacaggg tgataaagcc aagcaaagta
     2281 tcagggcaaa gtgtacagaa tatcttgata gagcagaaaa actaaaggag tacctgaaaa
     2341 ataaagagaa aaaagcacag aagccagtga aagaaggaca gccgagtcca gcagatgaga
     2401 aggggaatga cagtgatggg gaaggagaat ctgatgatcc tgaaaaaaag aaactacaga
     2461 atcaacttca aggtgccatt gttatagaac gaccaaatgt gaaatggagt gacgttgctg
     2521 gacttgaagg agccaaagaa gcactgaaag aggctgtgat actgcctatt aaatttcctc
     2581 atctttttac aggcaagaga acaccttgga ggggaatcct attatttggg ccgcctggaa
     2641 caggaaagtc ctacttagcc aaagctgtag caacagaagc caacaactca acattttttt
     2701 caatatcttc ctctgatctt gtttctaagt ggctaggtga aagtgaaaaa ctggttaaga
     2761 atttattcca acttgccaga gagaacaagc cctccattat cttcattgat gaaattgatt
     2821 ctctctgtgg ttcaagaagt gaaaatgaaa gtgaagccgc acgtagaatt aagacggagt
     2881 tcctagtgca aatgcaaggg gttggtgtag acaatgatgg aattttggtt ctgggagcta
     2941 caaatatacc ctgggttctg gattctgcca ttaggcgaag atttgagaaa cgaatttata
     3001 ttcccttgcc ggaaccccat gcccgagcag caatgtttaa actgcaccta gggaccactc
     3061 agaacagtct cacggaagca gactttcggg aacttgggag gaaaacagat ggttattcag
     3121 gggcagatat aagtatcatt gtacgtgatg cccttatgca gcctgttagg aaagtacagt
     3181 cagctactca ttttaaaaag gttcgcggac cttcccgagc tgatcctaac catcttgtag
     3241 atgatctgct aacaccttgc tctccaggtg accctggtgc cattgaaatg acatggatgg
     3301 atgtccctgg agataaactt ttggagccag ttgtttccat gtcggatatg ttgcggtcac
     3361 tatctaacac aaaacctaca gtcaatgaac atgacttgtt gaaattaaag aagtttacag
     3421 aagattttgg tcaagaaggc gtttaaacga attcgggctc ggtacccggg gatcctctag
     3481 agtcgacctg caggcatgca agctgatttg gctgctgcca ccgctgagca aggccgcttc
     3541 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     3601 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     3661 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     3721 tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga
     3781 attctaatat aacttcgtat agcatacatt atacgaagtt atggatccac tgttgacaat
     3841 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag
     3901 gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3961 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4021 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4081 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4141 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4201 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4261 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4321 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4381 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4441 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4501 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4561 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4621 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4681 tcgccttctt gacgagttct tctgattcga aatgaccgac caagcgacgc ccaaccggta
     4741 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4801 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4861 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4921 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4981 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5041 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5101 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5161 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5221 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5281 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5341 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5401 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct
     5461 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5521 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg aatcgtcaca
     5581 caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattgcggc catcgtgatg
     5641 gctagagttt agccctccca cacataacca gagggcagca attcacgaat cccaactgcc
     5701 gtcggctgtc catcactgtc cttcactatg gctttgatcc caggatgcag atcgagaagc
     5761 acctgtcggc accgtccgca ggggctcaag atgcccctgt tctcatttcc gatcgcgacg
     5821 atacaagtca ggttgccagc tgccgcagca gcagcagtgc ccagcaccac gagttctgca
     5881 caaggtcccc cagtaaaatg atatacattg acaccagtga agatgcggcc gtcgctagag
     5941 agagctgcgc tggcgacgct gtagtcttca gagatgggga tgctgttgat tgtagccgtt
     6001 gctctttcaa tgagggtgga ttcttcttga gacaaaggct tggccatggt ggcgctagtg
     6061 tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     6121 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     6181 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     6241 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     6301 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     6361 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     6421 ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat ttcaggccat
     6481 ggtgctgcgc aagacgcgtt aattaagacg t
//
