LOCUS       sequence-308960-        5674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses squirrel monkey CHMP3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5674)
  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 5674)
  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-squirrel-monkey-CHMP3
FEATURES             Location/Qualifiers
     source          1..5674
                     /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(2696..2715)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2761..2816
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2815..2834)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3177..3193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3177..3193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3190..3212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3201..3217
                     /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(3225..3231,3232..3249,3250..3255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3270..3291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3350..3545
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3396..3531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3551..3685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3588..3607)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3642..3661
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3753..3770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3924..4512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4004..4023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4683..5474,5475..5543))
                     /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     5306..5325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5544..5648)
                     /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 ggggsggggs 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 accatggggc tgtttggaaa aacccaggag aagccaccca aagaactggt caatgaatgg
     1981 tcattgaaga taagaaagga aatgagagtt gttgacaggc aaataaggga tatccaaaga
     2041 gaagaagaaa aagtgaaacg atctgtgaaa gatgcagcca agaagggcca gaaggatgtc
     2101 tgtgtagttc tggccaagga gatgatcagg tcaaggaagg ccgtgagcaa gctatatgca
     2161 tccaaagcgc acatgaattc ggtgctcatg gggatgaaga accagctcgc ggtcttgcga
     2221 gtggctggtt ccctgcagaa gagcacagaa gtgatgaagg ccatgcaaag tcttgtgaag
     2281 attccagaaa ttcaggccac catgagggag ctatccaaag aaatgatgaa ggctgggatc
     2341 atagaggaga tgttagagga cacttttgaa agcatggacg atcaggaaga aatggaggaa
     2401 gaagcagaaa tggaaattga caaaattcta tttgaaatta cagcaggggc cttgggcaaa
     2461 gcacccagta aagtgactga tgcccttcca gagccagaac cttcgggagc gatggctgcc
     2521 tcagaggatg aggaggagga ggaagaagct ctggaggcca tgcagtcccg gctggccaca
     2581 ctccgaagct gaatcttaat taattaactc gaggaattac tcctcaggtg caggctgcct
     2641 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     2701 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     2761 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     2821 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     2881 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     2941 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     3001 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     3061 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     3121 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     3181 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3241 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3301 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     3361 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3421 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3481 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3541 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3661 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     3721 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3781 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3841 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3901 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3961 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4021 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4081 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4141 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4201 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4261 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4321 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4381 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4441 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4501 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4561 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4621 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4681 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4741 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4801 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4861 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4921 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4981 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5041 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5101 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5161 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5221 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5281 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5341 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5401 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5461 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5521 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5581 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5641 gttccgcgca catttccccg aaaagtgcca cctg
//
