LOCUS       sequence-473357-        6490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag(R)-hVPS4A 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 6490)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6490)
  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            "Unpublished"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-hVPS4A Vector
FEATURES             Location/Qualifiers
     source          1..6490
                     /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"
     polyA_signal    3532..3653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3543..3562)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3768..3801)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3811..3858
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3890..4684
                     /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(3944..3963)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4554..4573
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4842..5430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4922..4941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5556..5604
                     /note="synthetic polyadenylation signal"
     CDS             complement(5628..6026)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6023..6032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6057..6414)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6071..6206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6223..6414
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6371..6391
                     /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 gacaacgtca accctccaga aagccattga tctggtgacg aaagccacag
     2161 aggaggacaa agccaagaac tacgaggagg cgctgcggct gtaccagcat gcggtggagt
     2221 acttcctcca cgctatcaag tatgaggccc acagcgacaa ggccaaggag agcattcgag
     2281 ccaagtgcgt gcagtaccta gaccgggccg agaagctgaa ggattattta cgaagcaaag
     2341 agaaacacgg caagaagcca gtcaaagaga accagagtga gggcaagggc agtgacagtg
     2401 acagtgaagg ggataatccg gagaaaaaga aactgcaaga acagctgatg ggtgccgtcg
     2461 tgatggagaa gcccaacata cggtggaacg acgtggccgg gctggagggg gccaaggagg
     2521 ccctcaaaga agctgtcatt ttgccaatca aattcccaca cttgttcaca ggcaagcgca
     2581 ccccctggcg ggggattctg ctgttcggac cccctggcac agggaaatcc tacctggcca
     2641 aagccgtggc aacagaggcc aacaactcca ccttcttctc tgtgtcctcc tcagatctga
     2701 tgtccaagtg gctgggggag agtgaaaagc tggtcaagaa cctgtttgag ctggccaggc
     2761 agcacaagcc ctccatcatc ttcatcgatg aggtggattc cctctgcggg tcccgaaatg
     2821 aaaatgagag tgaggccgcc cggaggatca aaacggagtt cttggtccag atgcaggggg
     2881 tggggaataa caatgatggg actctggttc ttggagccac aaacatccca tgggtgttgg
     2941 attcggccat caggaggagg tttgaaaaac gaatttatat ccccttgccg gaggaagctg
     3001 cccgcgccca gatgttccgg ttgcatctcg ggagcactcc ccacaacctc acggatgcaa
     3061 acatccacga gctggcccgg aagacggaag gctactcggg cgcggacatc agcatcatcg
     3121 tgcgggactc tctcatgcag cccgtgagga aggtgcagtc ggccacacac ttcaaaaagg
     3181 tctgtggccc ctctcgcacc aaccccagca tgatgattga tgacctcctg actccatgct
     3241 caccagggga cccaggagcc atggagatga cttggatgga tgtccctggg gacaaactct
     3301 tagagcctgt ggtttgcatg tcggacatgc tgcggtctct ggccaccacc cggcccacgg
     3361 tgaatgcaga cgacctcctg aaagtgaaga aattctcaga ggactttggg caagagagtg
     3421 tttaaacgaa ttcgggctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa
     3481 gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca
     3541 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3601 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3661 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     3721 agtaaaacct ctacaaatgt ggtaaaatct taattaagaa ttctaatata acttcgtata
     3781 gcatacatta tacgaagtta tggatccact gttgacaatt aatcatcggc atagtatatc
     3841 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca
     3901 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3961 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4021 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4081 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4141 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4201 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4261 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4321 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4381 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4441 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4501 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4561 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4621 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4681 ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta
     4741 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4801 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4861 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4921 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4981 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5041 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5101 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5161 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5221 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5281 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5341 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5401 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     5461 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5521 atcttcacct agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg
     5581 taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac
     5641 acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc
     5701 ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag
     5761 gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct
     5821 gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga
     5881 tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg
     5941 tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat
     6001 tcttcttgag acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg
     6061 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     6121 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     6181 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     6241 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     6301 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     6361 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta
     6421 cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgca agacgcgtta
     6481 attaagacgt
//
