LOCUS       sequence-328994-        5007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses ancestral New World monkey retroCHMP3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5007)
  AUTHORS   Rheinemann L, Downhour DM, Davenport KA, McKeown AN, Sundquist WI, 
            Elde NC
  TITLE     Recurrent evolution of an inhibitor of ESCRT-dependent virus budding
            and LINE-1 retrotransposition in primates.
  JOURNAL   Curr Biol. 2022 Feb 25. pii: S0960-9822(22)00240-8. doi: 
            10.1016/j.cub.2022.02.018.
  PUBMED    35245459
REFERENCE   2  (bases 1 to 5007)
  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: "Curr Biol. 
            2022 Feb 25. pii: S0960-9822(22)00240-8. doi: 
            10.1016/j.cub.2022.02.018."
COMMENT     Sequence Label: pEF1-alpha-ancestral-retroCHMP3-HA
FEATURES             Location/Qualifiers
     source          1..5007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(117..133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(117..133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(130..152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    141..157
                     /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(165..171,172..189,190..195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    210..231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        290..485
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      336..471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    491..625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(528..547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     582..601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(693..710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(864..1452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(944..963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1623..2414,2415..2483))
                     /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     2246..2265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2484..2588)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2650..3832
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2881..3823
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3780..3800
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4528..4554
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4643..4662)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4708..4763
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4762..4781)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
ORIGIN
        1 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
       61 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
      121 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      181 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
      301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      421 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      481 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
      541 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
      601 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
      661 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      721 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      781 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1261 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1441 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1501 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1561 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1621 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1681 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1741 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1801 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1861 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1921 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1981 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2041 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2101 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2161 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2221 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2281 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2341 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2401 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2461 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2521 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2581 gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat tgactagatc
     2641 atcgcgtgag gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     2701 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
     2761 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     2821 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     2881 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
     2941 ccttgaatta cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt
     3001 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
     3061 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
     3121 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
     3181 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
     3241 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
     3301 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     3361 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     3421 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     3481 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     3541 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
     3601 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     3661 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     3721 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     3781 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaattct
     3841 gcagtcgacg gtacccaatt tgccatgggg ctgtttggaa aaacccagga gaagccaccc
     3901 aaagaactgg tcaatgaatg gtcattgaag ataagaaagg aaatgagagt tgttgacagg
     3961 caaataaggg atatccaaag agaagaagaa aaagtgaaac gatctgtgaa agatgcagcc
     4021 aagaagggcc agaaggatgt ctgtgtagtt ctggccaagg agatgatcag gtcaaggaag
     4081 gccgtgagca agctgtatgc atccaaagca cacatgaact cggtgctcat ggggatgaag
     4141 aaccagctcg cggtcttgcg agtggctggt tccctgcaga agagcacaga agtgatgaag
     4201 gccatgcaaa gtcttgtgaa gattccagaa attcaggcca ccatgaggga gctgtccaaa
     4261 gaaatgatga aggctgggat catagaggag atgttagagg acacttttga aagcatggat
     4321 gatcaggaag aaatggagga agaagcagaa atggaaattg acaaaattct atttgaaatt
     4381 acagcagggg ccttgggcaa agcacccagt aaagtgactg atgcccttcc agagccagaa
     4441 ccttcaggag cgatggctgc ctcagaggat gaggaggagg aagaagctct ggaggccatg
     4501 cagtcccggc tggccacact ccgaagctac ccatacgatg ttccagatta cgcttgagcg
     4561 gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     4621 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     4681 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     4741 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     4801 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     4861 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     4921 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     4981 ttttttcttt aacatcccta aaatttt
//
