LOCUS       sequence-308882-        4809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses squirrel monkey retroCHMP3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4809)
  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 4809)
  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-squirrel-monkey-retroCHMP3-HA
FEATURES             Location/Qualifiers
     source          1..4809
                     /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             1680..1706
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1795..1814)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1860..1915
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1914..1933)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2276..2292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2276..2292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2289..2311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2300..2316
                     /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(2324..2330,2331..2348,2349..2354))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2369..2390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2449..2644
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2495..2630
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2650..2784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2687..2706)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2741..2760
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2852..2869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3023..3611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3103..3122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3782..4573,4574..4642))
                     /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     4405..4424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4643..4747)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4809,1..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 gtacccaatt tgccatggga ctcttggtta aaacccagga gaagccacca aaagaactgg
     1261 tcaatgaatg gtcattgaag ataagaaagg aaatgagagt tgttgacagg caaatacggg
     1321 atatccaaag agaagaagaa aaagtgaaac gatctgtgaa agaagcagtc aagaagggcc
     1381 agaaggatgt ctgcacagtt ctggccaggg agatgatccg gtcagggacg gctgtgagca
     1441 agctgtatga atccaaagcg cacctggact cggtgctcac ggggatgagg aaccagttgg
     1501 cggtcttgcg aggggctggt tccctgcaga agagtgcaga agtgacgagg gctgtgcaaa
     1561 gtcttgtgaa ggttccagaa attcaggcca ccatgaggga gctgtccaaa gaaatgacga
     1621 aggctgggat catagaggag atgttagagg acactttcga aagcatggac gatcgggaat
     1681 acccatacga tgttccagat tacgcttgag cggccgcact cctcaggtgc aggctgccta
     1741 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     1801 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     1861 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     1921 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     1981 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     2041 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     2101 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     2161 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     2221 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     2281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2341 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     2461 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2521 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2581 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2641 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2701 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2761 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     2821 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2881 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2941 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3781 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3841 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3901 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3961 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4021 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4081 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4141 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4201 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4261 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4321 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4381 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4441 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4501 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4561 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4621 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4681 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4741 ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt gactagatca
     4801 tcgcgtgag
//
