LOCUS       sequence-308929-        5459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human CHMP3(150) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5459)
  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 5459)
  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: pCAG-OSF-PP-human-CHMP3(150)
FEATURES             Location/Qualifiers
     source          1..5459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1753..1762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1768..1791
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1828..1851
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1858..1881
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1894..1917
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2481..2500)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2546..2601
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2600..2619)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2962..2978)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2962..2978)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2975..2997)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2986..3002
                     /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(3010..3016,3017..3034,3035..3040))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3055..3076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3135..3330
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3181..3316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3336..3470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3373..3392)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3427..3446
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3538..3555)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3709..4297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3789..3808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4468..5259,5260..5328))
                     /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     5091..5110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5329..5433)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccgggggg gggggggggg gggggggggg
      541 gggggggggs grggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgatc
     1741 gacggtatcg atgccaccat ggctagctgg agccacccgc agttcgagaa aggtggaggt
     1801 gcccgaggtg gatcgggagg tggatcgtgg agccacccgc agttcgaaaa aggattcgat
     1861 tacaaggatg acgatgacaa gggcacaaga tctctggaag tcctgttcca gggcccgggt
     1921 accatgggcc tgtttggcaa gacccaggag aagccgccca aagaactggt caatgagtgg
     1981 tcattgaaga taagaaagga aatgagagtt gttgacaggc aaataaggga tatccaaaga
     2041 gaagaagaaa aagtgaaacg atctgtgaaa gatgctgcca agaagggcca gaaggatgtc
     2101 tgcatagttc tggccaagga gatgatcagg tcaaggaagg ctgtgagcaa gctgtatgca
     2161 tccaaagcac acatgaactc agtgctcatg gggatgaaga accagctcgc ggtcttgcga
     2221 gtggctggtt ccctgcagaa gagcacagaa gtgatgaagg ccatgcaaag tcttgtgaag
     2281 attccagaga ttcaggccac catgagggag ttgtccaaag aaatgatgaa ggctgggatc
     2341 atagaggaga tgttagagga cacttttgaa agctgaatct taattaatta actcgaggaa
     2401 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     2461 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     2521 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     2581 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     2641 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     2701 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     2761 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     2821 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     2881 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     2941 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3001 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3061 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3121 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3241 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3301 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     3361 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3421 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     3481 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3541 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3601 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3661 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3721 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3781 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3841 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3901 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3961 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4021 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4081 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4141 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4201 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4261 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4321 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4381 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4441 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4501 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4561 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4621 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4681 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4741 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4801 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4861 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4921 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4981 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5041 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5101 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5161 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5221 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5281 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5341 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
