LOCUS       sequence-298385-        4781 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 4781)
  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 4781)
  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..4781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..1217
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          266..1208
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1165..1185
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1690..1716
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1805..1824)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1870..1925
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1924..1943)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2284..2300)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2284..2300)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2297..2319)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2308..2324
                     /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(2332..2338,2339..2356,2357..2362))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2377..2398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2457..2652
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2503..2638
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2658..2792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2695..2714)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2749..2768
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2860..2877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3031..3619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3111..3130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3790..4581,4582..4650))
                     /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     4413..4432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4651..4755)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agatcatcgc gtgaggctcc ggtgcccgtc agtgggcaga
       61 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
      121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
      181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
      241 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
      301 ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg ctgcagtacg
      361 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
      421 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
      481 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
      541 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
      601 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
      661 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
      721 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
      781 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
      841 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
      901 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
      961 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     1021 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     1081 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     1141 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     1201 catttcaggt gtcgtgagga attctgcagt cgacggtacc aatttgccat gggactcttt
     1261 ggaaaaaccc aggagaagcc tcccaaagag ctggttaatg aatggtcact gaagatcaga
     1321 aaggaaatga gagttgttga caggcaaata agagatatcc aaagagaaga agagaaagta
     1381 aaacggtctg tgaaagatgc agccaagaag ggccaaaagg aagtctgtgt ggttctggcc
     1441 aaggagatga tcaggtcaag gaaagccgtg agcaagctct atgcctccaa agcacacatg
     1501 aactctgtgc tcatgggcat gaagaaccaa ctcgcggtcc tgcgagtagc tggttctctg
     1561 cagaagagca cagaagtgat gaaggcaatg cagagtcttg tgaagatccc agaaatccag
     1621 gccaccatgc gggagctgtc caaagagatg atgaaggctg gaatcataga ggagatgtta
     1681 gaggatacat acccatacga tgttccagat tacgcttgag cggccgcact cctcaggtgc
     1741 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     1801 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     1861 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     1921 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     1981 tttagagttt ggcaacatat gccatatgct ggctgccatg aacaaaggtg gctataaaga
     2041 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     2101 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     2161 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     2221 atagctgtcc ctcttctctt atgaagatcc ctcgacctgc agcccaagct tggcgtaatc
     2281 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2341 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2401 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     2461 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2521 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2581 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2641 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2701 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2761 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     2821 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2881 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2941 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3001 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3061 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     3241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3361 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3421 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3481 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3541 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3601 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3661 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3721 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3781 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3841 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3901 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3961 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4021 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4081 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4141 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4201 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4261 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4321 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4381 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4441 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4501 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4561 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4621 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4681 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4741 aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
