LOCUS       sequence-308917-        4784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mouse CHMP3(147) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4784)
  AUTHORS   Rheinemann L, Downhour DM, Bredbenner K, Mercenne G, Davenport KA, 
            Schmitt PT, Necessary CR, McCullough J, Schmitt AP, Simon SM, 
            Sundquist WI, Elde NC
  TITLE     RetroCHMP3 blocks budding of enveloped viruses without blocking 
            cytokinesis.
  JOURNAL   Cell. 2021 Sep 27. pii: S0092-8674(21)01055-2. doi: 
            10.1016/j.cell.2021.09.008.
  PUBMED    34597582
REFERENCE   2  (bases 1 to 4784)
  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: "Cell. 2021 
            Sep 27. pii: S0092-8674(21)01055-2. doi: 
            10.1016/j.cell.2021.09.008."
COMMENT     Sequence Label: pEF1-alpha-mouse-CHMP3(147)-HA
FEATURES             Location/Qualifiers
     source          1..4784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          231..1173
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1655..1681
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1770..1789)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1835..1890
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1889..1908)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2251..2267)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2251..2267)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2264..2286)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2275..2291
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2299..2305,2306..2323,2324..2329))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2344..2365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2424..2619
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2470..2605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2625..2759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2662..2681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2716..2735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2827..2844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2998..3586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3078..3097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3757..4548,4549..4617))
                     /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     4380..4399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4618..4722)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4784,1..1182)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaattct gcagtcgacg
     1201 gtaccaattt gccatgggac tctttggaaa aacccaggag aagcctccca aagagctggt
     1261 taatgaatgg tcactgaaga tcagaaagga aatgagagtt gttgacaggc aaataagaga
     1321 tatccaaaga gaagaagaga aagtaaaacg gtctgtgaaa gatgcagcca agaagggcca
     1381 aaaggaagtc tgtgtggttc tggccaagga gatgatcagg tcaaggaaag ccgtgagcaa
     1441 gctctatgcc tccaaagcac acatgaactc tgtgctcatg ggcatgaaga accaactcgc
     1501 ggtcctgcga gtagctggtt ctctgcagaa gagcacagaa gtgatgaagg caatgcagag
     1561 tcttgtgaag atcccagaaa tccaggccac catgcgggag ctgtccaaag agatgatgaa
     1621 ggctggaatc atagaggaga tgttagagga tacataccca tacgatgttc cagattacgc
     1681 ttgagcggcc gcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     1741 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     1801 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     1861 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     1921 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     1981 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     2041 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     2101 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     2161 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
     2221 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2281 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2341 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2401 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     2461 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2521 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2581 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     2641 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2701 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     2761 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2821 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2941 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3181 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3361 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3421 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3481 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3541 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3601 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3721 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3781 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3841 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3901 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3961 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4021 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4081 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4141 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4201 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4261 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4321 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4381 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4441 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4501 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4561 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4621 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4681 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4741 gccacctggg tcgacattga ttattgacta gatcatcgcg tgag
//
