LOCUS       sequence-329050-        4809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses reconstructed New World monkey retroCHMP3 Node 3 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, 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 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: "Curr Biol. 
            2022 Feb 25. pii: S0960-9822(22)00240-8. doi: 
            10.1016/j.cub.2022.02.018."
COMMENT     Sequence Label: pEF1-alpha -NWM-Node 3
FEATURES             Location/Qualifiers
     source          1..4809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(229..245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(229..245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(242..264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    253..269
                     /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(277..283,284..301,302..307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    322..343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        402..597
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      448..583
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    603..737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(640..659)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     694..713
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(805..822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(976..1564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1056..1075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1735..2526,2527..2595))
                     /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     2358..2377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2596..2700)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2762..3944
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2993..3935
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3892..3912
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4442..4468
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4557..4576)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4622..4677
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4676..4695)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
ORIGIN
        1 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
       61 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
      121 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
      181 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
      241 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      301 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
      361 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
      421 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
      481 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
      541 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
      601 ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      661 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      721 cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      781 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      841 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      901 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      961 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1381 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1441 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1501 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1561 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1621 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1681 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1741 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1801 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1861 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1921 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1981 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2041 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2101 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2161 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2221 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2281 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2341 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2401 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2461 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2521 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2581 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2641 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2701 cacatttccc cgaaaagtgc cacctgggtc gacattgatt attgactaga tcatcgcgtg
     2761 aggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     2821 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     2881 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     2941 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     3001 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     3061 tacttccacg cccctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
     3121 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
     3181 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
     3241 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
     3301 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
     3361 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
     3421 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
     3481 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
     3541 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
     3601 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
     3661 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
     3721 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     3781 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
     3841 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     3901 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt ctgcagtcga
     3961 cggtacccaa tttgccatgg ggctgttggt taaaacccag gagaagccac caaaagaact
     4021 ggtcaatgaa tggtcattga agataagaaa ggaaatgaga gttgttgaca ggcaaataag
     4081 ggatatccaa agagaagaag aaaaagtgaa acgatctgtg aaagatgcag tcaagaaggg
     4141 ccagaaggat gtctgcacag ttctggccag ggagatgatc aggtcaggga cggccgtgag
     4201 caagctgtat gcatccaaag cgcacatgaa ctcggtgctc atggggatga ggaaccagct
     4261 cgcggtcttg cgagtggctg gttccctgca gaagagcaca gaagtgatga gggccgtgca
     4321 aagtcttgtg aagattccag aaattcaggc caccatgagg gagctgtcca aagaaatgat
     4381 gaaggctggg atcatagagg agatgttaga ggacactttc gaaagcatgg acgatcagga
     4441 atacccatac gatgttccag attacgcttg agcggccgca ctcctcaggt gcaggctgcc
     4501 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     4561 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     4621 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     4681 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     4741 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     4801 cagtatatg
//
