LOCUS       sequence-407398-        7731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Onyong-nyong virus E1E2 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7731)
  AUTHORS   Strebinger D, Frangieh CJ, Friedrich MJ, Faure G, Macrae RK, Zhang F
  TITLE     Cell type-specific delivery by modular envelope design.
  JOURNAL   Nat Commun. 2023 Aug 23;14(1):5141. doi: 10.1038/s41467-023-40788-8.
  PUBMED    37612276
REFERENCE   2  (bases 1 to 7731)
  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: "Nat 
            Commun."; date: "2023-08-23"; volume: "14(1):5141. doi"; pages: " 
            10.1038/s41467-023-40788-8"
COMMENT     Sequence Label: pHCMV_ONNV_E1E2
FEATURES             Location/Qualifiers
     source          1..7731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          792..1364
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      1419..1428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4436..4455)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4501..4556
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4555..4574)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4914..4930)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4914..4930)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4927..4949)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4938..4954
                     /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(4962..4968,4969..4986,4987..4992))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5007..5028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5145..5162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5316..5904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5396..5415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6075..6866,6867..6935))
                     /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     6698..6717
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6936..7040)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7066..7521)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7203..7224)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7415..7434
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7648..7670
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7662..7679
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7663..7679
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7689..7708
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7689..7707
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca
       61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaca cgccatccac
      721 gctgttttga cctccataga tgacaccggg accgatccag cctccggtcg accgatcctg
      781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      961 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
     1021 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
     1081 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1141 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1201 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt
     1261 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1321 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc
     1381 tggttgttgt gctgtctcat cattttggca aagaattcgc caccatggga agcctcgccc
     1441 tccccgtgat gtgtctgctg gccaatacaa cctttccttg ctcccaaccc ccttgcgccc
     1501 cttgctgtta cgagaagaaa cccgaggaga cactgaggat gctcgaggac aatgtgatgc
     1561 agcccggcta ttaccaactg ctggatagcg ccctcgcttg cagccaaagg agacaaaaga
     1621 ggaacgctag ggagaacttt aacgtctata aagtgacaag gccttatctg gctcactgtc
     1681 ccgactgtgg cgaaggccat agctgccatt cccctatcgc tctggaaaga attaggagcg
     1741 aagccacaga cggcacactg aagatccaag tgtctctgca gatcggcatc aagacagacg
     1801 actcccacga ttggacaaag ctgagataca tggacagcca tacccccgtg gatgctgaca
     1861 gatccggact gttcgtgaga acatccgctc cttgcaccat taccggcacc atgggacatt
     1921 tcatcctcgc tagatgccct aagggcgaaa cactcaccgt cggatttgtg gactctagaa
     1981 gaatctccca cacatgtatg caccccttca gacatgagcc ccccctcatc ggaagagaga
     2041 agttccactc tagacctcaa cacggcaaag agctgccttg ctccacatac gtgcacacca
     2101 ccgctgccac cgccgaagag attgaggtgc acatgcctcc cgataccccc gattatacac
     2161 tgatgaccca acaagccgga aacgtgaaga tcaccgtgga cggccagaca gtgagataca
     2221 agtgcaagtg tgacggctcc aacgagggac tcatcaccgc tgataaggtg attaataact
     2281 gtaaagtcga ccaatgtcac accgccgtca ccaaccacaa gaagtggcag tacaactccc
     2341 ctctgacccc tagaaacagc gaacaaggcg acagaaaggg aaagatccac atccctttcc
     2401 ctctcgtgaa caccacatgc agagtcccta aggctaggaa ccccaccgtc acctacggaa
     2461 aaaatagggt gacactgctg ctgcaccccg accaccctac cctcctcagc tacagagcta
     2521 tgggaagaat ccccgactac catgaagagt ggatcacaaa taagaaagag atcagcatta
     2581 cagtgcccgc cgagggactc gaagtgacat ggggcaacaa cgacccctac aaatactggc
     2641 cccaactgag caccaacgga acagctcacg gccaccccca cgaaatcatt ctgtactact
     2701 acgagctgta ccccaccaca acaatcgctg tgctcgccgc cgcctccatc gtcattacct
     2761 ctctggtcgg actgtccctc ggcatgtgta tctgcgctag aagaagatgc atcacccctt
     2821 acgaactcac acccggcgcc accattccct ttctgctggg agtgctgtgc tgcgctagaa
     2881 cagccaaggc cgcctcctac tatgaggccg ccacatatct ctggaatgaa cagcagcccc
     2941 tcttctggct gcagctgctg attcctctgt ccgctgccat cgtggtgtgc aactgcctca
     3001 agctgctgcc ttgttgttgc aagacactga cattcctcgc cgtgatgagc attggagcta
     3061 ggaccgtcac cgcctacgaa cacgctaccg tgatccccaa taccgtcggc gtgccttgca
     3121 aaacactggt gagcagaccc ggctacagcc ctatggtgct ggaaatggag ctgcagtccg
     3181 tgacactgga acccgctctg tccctcgact acatcacatg cgagtacaaa accatcaccc
     3241 ccagccccta cgtgaagtgt tgcggcaccg ctgaatgcaa agccaagaat ctccccgact
     3301 ataactgcaa ggtgtttacc ggcgtgtacc ctttcatgtg gggcggagcc tattgcttct
     3361 gcgacgccga aaacacacaa ctgtccgaag ctcatgtcga gaagagcgaa agctgcaaaa
     3421 ccgagttcgc ctccgcctat agggctcata cagccagcgt gagcgctaag ctgagagtgt
     3481 tctaccaagg caacaacatt acagtgagcg cttacgccaa cggagatcac gccgtgacag
     3541 tggaggacgc taaattcgtg attggccctc tgagcagcgc ttggtcccct ttcgacaaca
     3601 agatcgtggt gtacaaggga gaggtgtaca acatggacta tccccccttt ggagctggaa
     3661 ggcccggcca gttcggagat atccagtcta gaacacccga ctccaaggac gtgtacgcta
     3721 acacccagct gattctccaa aggcccgccg ctggcgccat ccatgtgccc tatagccaag
     3781 cccctagcgg ctttaagtac tggctcaagg aaaagggcgc ttctctgcag cacacagctc
     3841 ccttcggctg ccagatcgct acaaaccccg tcagagccgt gaattgcgcc gtcggcaata
     3901 tcccagtgag cattgacatc cccgacgccg cttttacaag agtcacagat gccccctcca
     3961 tcaccgacat gagctgcgag gtggctagct gcacacacag ctccgatttt ggcggcgctg
     4021 ccgtcatcaa gtacaccgct tccaagaagg gcaagtgcgc cgtccactcc gtcaccaacg
     4081 ctgtcaccat tagagagcct aatgtggatg tgaagggcac agcccagctg caaatcgcct
     4141 tcagcacagc tctggcctcc gctgagttta aggtccaaat ctgctccaca ctggtgcatt
     4201 gcagcgctac atgccacccc cccaaggacc atattgtcaa ctatcccagc ccccacacaa
     4261 ccctcggagt gcaagacatc agcacaaccg ccatgagctg ggtgcaaaag attaccggcg
     4321 gagtgggact ggtggtcgcc atcgctgctc tcatcctcat catcgtgctc tgtgtgagct
     4381 ttagcagaca ctgatgtctt cctggagctc cagtcattga ctataacgtc tcagatcttt
     4441 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     4501 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     4561 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     4621 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     4681 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     4741 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     4801 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     4861 tccctcttct cttatggaga tccctcgacg gatcggccgc aattcgtaat catgtcatag
     4921 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4981 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5041 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5101 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6001 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6061 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6121 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6181 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggcaccagat
     6241 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6301 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6361 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6421 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6481 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6541 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6601 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6661 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6721 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6781 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6841 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6901 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6961 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7021 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     7081 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     7141 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     7201 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     7261 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     7321 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     7381 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     7441 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     7501 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     7561 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     7621 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     7681 agcgcgcgta atacgactca ctatagggcg aattggagct ccaccgcggt g
//
