LOCUS       sequence-329049-        4809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses reconstructed New World monkey retroCHMP3 Node 2 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 2
FEATURES             Location/Qualifiers
     source          1..4809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    106..240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(143..162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     197..216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(308..325)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(479..1067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(559..578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1238..2029,2030..2098))
                     /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     1861..1880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2099..2203)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2265..3447
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2496..3438
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3395..3415
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3945..3971
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4060..4079)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4125..4180
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4179..4198)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4541..4557)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4541..4557)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4554..4576)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4565..4581
                     /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(4589..4595,4596..4613,4614..4619))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4634..4655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4714..4809,1..100)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(4760..4809,1..86)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
       61 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
      121 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
      181 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
      241 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
      301 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      361 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
      421 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      481 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      541 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      601 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      661 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      721 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      781 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      841 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      901 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      961 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1021 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1081 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1141 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1201 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1261 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1321 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1381 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1441 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1501 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1561 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1621 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1681 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1741 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1801 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1861 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1921 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1981 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2041 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2101 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2161 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2221 tgccacctgg gtcgacattg attattgact agatcatcgc gtgaggctcc ggtgcccgtc
     2281 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     2341 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     2401 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     2461 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     2521 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg
     2581 ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg
     2641 ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
     2701 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     2761 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     2821 gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     2881 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     2941 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     3001 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     3061 gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg
     3121 ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc
     3181 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc
     3241 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     3301 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     3361 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     3421 ttttttcttc catttcaggt gtcgtgagga attctgcagt cgacggtacc caatttgcca
     3481 tggggctgtt ggttaaaacc caggagaagc caccaaaaga actggtcaat gaatggtcat
     3541 tgaagataag aaaggaaatg agagttgttg acaggcaaat aagggatatc caaagagaag
     3601 aagaaaaagt gaaacgatct gtgaaagatg cagccaagaa gggccagaag gatgtctgca
     3661 cagttctggc caaggagatg atcaggtcaa ggaaggccgt gagcaagctg tatgcatcca
     3721 aagcgcccat gaactcggtg ctcatgggga tgaggaacca gctcgcggtc ttgcgagtgg
     3781 ctggttccct gcagaagagc acagaagtga tgagggccgt gcaaagtctt gtgaagattc
     3841 cagaaattca ggccaccatg agggagctgt ccaaagaaat gatgaaggct gggatcatag
     3901 aggagatgtt agaggacact ttcgaaagca tggacgatca ggaataccca tacgatgttc
     3961 cagattacgc ttgagcggcc gcactcctca ggtgcaggct gcctatcaga aggtggtggc
     4021 tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa
     4081 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     4141 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     4201 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     4261 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     4321 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     4381 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     4441 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     4501 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4561 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4621 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4681 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     4741 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4801 ccatggctg
//
