LOCUS       sequence-472424-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HiBit-VPS4B in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6633)
  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            "Unpublished"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: HiBit-VPS4B Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3..124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14..33)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     68..87
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        369..726
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        369..560
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(392..412)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      577..712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             777..1571
                     /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(831..850)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1441..1460
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1635..1683
                     /note="synthetic polyadenylation signal"
     promoter        1839..1943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1944..2012,2013..2804)
                     /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(2162..2181)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2962..3550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3042..3061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3666..3949
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        4140..4519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4520..4723
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4674..4694
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          4859..4991
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4974..4993
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     5035..5054
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5035..5053
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5959..5970
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(6581..6599)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
       61 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      121 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
      181 ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc gaattttaac aaaatattaa
      241 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      301 catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
      361 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
      421 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      481 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      541 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      601 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      661 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
      721 tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taaggctaga gccaccatga
      781 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
      841 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
      901 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
      961 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     1021 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     1081 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     1141 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     1201 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     1261 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     1321 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     1381 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     1441 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     1501 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     1561 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     1621 atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     1681 gtgaatcgat agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc
     1741 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg
     1801 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1861 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1921 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1981 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2041 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2101 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc
     2161 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2221 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2281 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2341 ccaacttact tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2401 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2461 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2521 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2581 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2641 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2701 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2761 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc
     2821 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2881 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3001 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3061 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3121 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag
     3181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3361 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3481 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3541 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3661 atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg
     3721 aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat
     3781 aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt
     3841 tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa
     3901 ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca
     3961 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat cttcaatatt ggccattagc
     4021 catattattc attggttata tagcataaat caatattggc tattggccat tgcatacgtt
     4081 gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg
     4141 gcattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     4201 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     4261 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     4321 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     4381 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     4441 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
     4501 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
     4561 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     4621 gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat
     4681 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     4741 gatcactaga agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc
     4801 ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt
     4861 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     4921 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     4981 tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga gtattaatac
     5041 gactcactat agggctagcg ctcaccatgg tgagcggctg gcggctgttc aagaagatta
     5101 gcgggagctc cggtggctcg agcggtgcga tcgccatgtc atccacttcg cccaacctcc
     5161 agaaagcgat agatctggct agcaaagcag cgcaagaaga caaggctggg aactacgaag
     5221 aagcccttca gctctatcag catgctgtgc agtattttct tcatgtcgtt aaatatgaag
     5281 cacagggtga taaagccaag caaagtatca gggcaaagtg tacagaatat cttgatagag
     5341 cagaaaaact aaaggagtac ctgaaaaata aagagaaaaa agcacagaag ccagtgaaag
     5401 aaggacagcc gagtccagca gatgagaagg ggaatgacag tgatggggaa ggagaatctg
     5461 atgatcctga aaaaaagaaa ctacagaatc aacttcaagg tgccattgtt atagaacgac
     5521 caaatgtgaa atggagtgac gttgctggac ttgaaggagc caaagaagca ctgaaagagg
     5581 ctgtgatact gcctattaaa tttcctcatc tttttacagg caagagaaca ccttggaggg
     5641 gaatcctatt atttgggccg cctggaacag gaaagtccta cttagccaaa gctgtagcaa
     5701 cagaagccaa caactcaaca tttttttcaa tatcttcctc tgatcttgtt tctaagtggc
     5761 taggtgaaag tgaaaaactg gttaagaatt tattccaact tgccagagag aacaagccct
     5821 ccattatctt cattgatgaa attgattctc tctgtggttc aagaagtgaa aatgaaagtg
     5881 aagccgcacg tagaattaag acggagttcc tagtgcaaat gcaaggggtt ggtgtagaca
     5941 atgatggaat tttggttctg ggagctacaa atataccctg ggttctggat tctgccatta
     6001 ggcgaagatt tgagaaacga atttatattc ccttgccgga accccatgcc cgagcagcaa
     6061 tgtttaaact gcacctaggg accactcaga acagtctcac ggaagcagac tttcgggaac
     6121 ttgggaggaa aacagatggt tattcagggg cagatataag tatcattgta cgtgatgccc
     6181 ttatgcagcc tgttaggaaa gtacagtcag ctactcattt taaaaaggtt cgcggacctt
     6241 cccgagctga tcctaaccat cttgtagatg atctgctaac accttgctct ccaggtgacc
     6301 ctggtgccat tgaaatgaca tggatggatg tccctggaga taaacttttg gagccagttg
     6361 tttccatgtc ggatatgttg cggtcactat ctaacacaaa acctacagtc aatgaacatg
     6421 acttgttgaa attaaagaag tttacagaag attttggtca agaaggcgtt taaacgaatt
     6481 cgggctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc tgatccggct
     6541 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     6601 taaccccttg gggcggccgc ttcgagcaga cat
//
