LOCUS       sequence-406228-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Chikungunya virus E1E2 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  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 8508)
  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_CHIKV_E1E2
FEATURES             Location/Qualifiers
     source          1..8508
                     /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(5213..5232)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5278..5333
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5332..5351)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5691..5707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5691..5707)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5704..5726)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5715..5731
                     /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(5739..5745,5746..5763,5764..5769))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5784..5805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5922..5939)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6093..6681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6173..6192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6852..7643,7644..7712))
                     /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     7475..7494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7713..7817)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7843..8298)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7980..8001)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8192..8211
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8425..8447
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8439..8456
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8440..8456
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8466..8485
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8466..8484
                     /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 caccatggag tttatcccta
     1441 cacaaacatt ttataacaga aggtatcagc ctagaccttg gacccccaga cctacaattc
     1501 aagtgattag acctaggcct agacctcaaa ggcaagccgg acagctcgcc cagctcatct
     1561 ccgctgtcaa caaactgacc atgagggccg tcccccaaca gaagcctaga aaaaacagaa
     1621 agaacaaaaa acagaaacag aagcagcaag ccccccagaa taacacaaat cagaagaagc
     1681 agccccctaa aaagaagccc gcccagaaga agaagaagcc cggaagaaga gagagaatgt
     1741 gtatgaaaat cgaaaatgac tgcattttcg aggtcaaaca cgaaggcaag gtgaccggct
     1801 acgcttgtct ggtgggcgac aaggtgatga agcccgccca cgtgaagggc acaatcgaca
     1861 acgccgatct ggccaagctg gcctttaaga ggtccagcaa gtacgacctc gaatgtgccc
     1921 agatccccgt gcacatgaag tccgatgcca gcaagttcac acacgagaag cccgaaggct
     1981 actataactg gcaccatgga gccgtgcagt atagcggagg aagatttacc attcccaccg
     2041 gagccggaaa acccggcgac agcggaaggc ccattttcga caacaagggc agagtggtcg
     2101 ccatcgtgct cggaggcgct aacgaaggag ctagaacagc tctctccgtg gtgacatgga
     2161 acaaagacat cgtcaccaag atcacacccg agggagctga ggaatggagc ctcgccattc
     2221 ccgtgatgtg tctgctggct aacacaacat tcccttgcag ccagcctcct tgcattcctt
     2281 gttgctacga gaaggagccc gaagagacac tgaggatgct cgaggataac gtcatgagac
     2341 ccggctacta tcagctgctg caagcctctc tgacatgctc cccccatagg cagagaagat
     2401 ccaccaagga taactttaac gtgtataagg ccacaagacc ctacctcgct cattgccccg
     2461 actgcggcga gggccatagc tgtcacagcc ccgtggctct ggaaaggatt aggaacgagg
     2521 ctaccgatgg aacactgaag atccaagtct ctctgcagat cggcattggc acagacgatt
     2581 cccacgattg gacaaagctg agatacatgg acaatcacat ccccgccgat gccggaagag
     2641 ccggactctt tgtgaggaca agcgctcctt gcaccatcac cggcacaatg ggccatttta
     2701 tcctcgccag atgtcccaaa ggcgagacac tgacagtcgg cttcaccgat tccagaaaga
     2761 ttagccacag ctgcacccac cctttccatc atgacccccc cgtgattgga agggagaagt
     2821 tccatagcag acctcaacac ggaaaggaac tcccttgctc cacctatgtg caaagcaatg
     2881 ccgccaccgc cgaagagatc gaggtgcata tgccccccga cacccccgac agaacactgc
     2941 tgagccaaca atccggcaac gtgaagatca cagtcaacag ccagaccgtg aggtacaagt
     3001 gcaactgcgg aggctccaat gagggcctca ttacaaccga caaggtcatc aacaactgca
     3061 aggtcgacca gtgccacgct gctgtcacca accataagaa atggcagtat aacagccctc
     3121 tggtgcctag aaatgccgag ctgggcgata gaaagggcaa gatccatatt cccttccctc
     3181 tggccaacgt cacatgtatg gtgcccaagg ccagaaaccc tacagtcacc tacggcaaaa
     3241 atcaagtgat catgctgctc taccccgatc atcctacact gctgtcctat agaagcatgg
     3301 gagaagagcc caattaccaa gaggagtggg tgacccacaa aaaggaggtc gtcctcaccg
     3361 tgcctacaga gggcctcgag gtgacatggg gcaataacga gccctacaag tactggcccc
     3421 agctgtccgc taacggaacc gcccacggcc acccccatga gattattctg tactattacg
     3481 agctgtaccc caccatgacc gtggtggtcg tgtccgtcgc tagctttatt ctgctgagca
     3541 tggtgggaat ggccgtcgga atgtgcatgt gtgctagaag gaggtgcatt acaccctatg
     3601 aactgacacc cggagctacc gtcccctttc tgctgtctct gatctgctgt attagaacag
     3661 ccaaagccgc cacatatcaa gaagccgccg tctacctctg gaatgagcag cagcccctct
     3721 tttggctgca agctctgatt cccctcgccg ctctcatcgt gctgtgcaat tgtctgagac
     3781 tgctgccttg ctgctgcaaa accctcgcct ttctggctgt catgagcatc ggcgctcaca
     3841 ccgtgagcgc ttacgagcac gtcaccgtga tccccaacac cgtgggcgtc ccctacaaga
     3901 ccctcgtgaa tagacccggc tacagcccta tggtgctgga aatggagctg ctgtccgtca
     3961 cactggaacc cacactgtcc ctcgactaca tcacatgcga gtacaagacc gtcatccctt
     4021 ccccttacgt gaagtgctgc ggcaccgccg agtgcaagga caaaaatctc cccgactaca
     4081 gctgcaaggt gttcaccggc gtgtatccct ttatgtgggg aggcgcctat tgcttctgcg
     4141 acgccgagaa tacccaactg tccgaggctc atgtcgagaa atccgagagc tgcaaaaccg
     4201 agttcgccag cgcctacaga gcccacacag ctagcgcttc cgctaagctg agagtgctgt
     4261 accaaggcaa taacatcacc gtgaccgctt atgccaacgg agatcatgcc gtcacagtga
     4321 aggacgccaa attcattgtg ggccccatga gctccgcttg gacacccttc gataacaaga
     4381 tcgtcgtgta caagggcgac gtctacaaca tggattaccc cccttttgga gctggcagac
     4441 ccggccaatt cggagatatt caatctagaa cccccgaaag caaggatgtg tacgccaaca
     4501 cccaactcgt gctgcaaagg cccgccgccg gaacagtcca tgtcccctat agccaagctc
     4561 ctagcggctt caagtactgg ctgaaagaga ggggagcttc tctgcagcat acagctcctt
     4621 tcggatgcca gatcgccacc aatcccgtga gggccatgaa ttgtgccgtc ggcaacatgc
     4681 ccatttccat cgatatcccc gatgccgctt tcacaagggt ggtggatgcc ccctctctga
     4741 cagacatgag ctgtgaagtg cccgcttgta cccacagcag cgacttcggc ggagtcgcca
     4801 ttattaagta cgccgtgagc aaaaagggca agtgtgccgt gcacagcatg accaatgccg
     4861 tgaccattag agaggccgag atcgaagtcg aaggcaattc ccagctccaa atcagcttta
     4921 gcaccgccct cgcctccgct gagttcagag tccaagtgtg cagcacacaa gtgcactgtg
     4981 ctgctgagtg tcatcccccc aaagaccaca tcgtcaacta ccccgctagc cacacaacac
     5041 tgggcgtgca agacatctcc gctacagcca tgagctgggt ccaaaagatc accggaggag
     5101 tgggactcgt cgtcgctgtc gccgctctga ttctgatcgt ggtgctctgt gtgtccttct
     5161 ctagacattg atgtcttcct ggagctccag tcattgacta taacgtctca gatctttttc
     5221 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     5281 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     5341 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     5401 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     5461 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     5521 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     5581 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     5641 ctcttctctt atggagatcc ctcgacggat cggccgcaat tcgtaatcat gtcatagctg
     5701 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5761 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5821 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5881 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5941 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6001 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6061 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6121 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6181 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6241 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6301 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6361 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6421 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6481 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6541 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6601 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6661 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6721 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6781 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6841 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6901 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6961 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc accagattta
     7021 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7081 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7141 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7201 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7261 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7321 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7381 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7441 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7501 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7561 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7621 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7681 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7741 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7801 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     7861 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     7921 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     7981 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     8041 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     8101 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     8161 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     8221 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     8281 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8341 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8401 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     8461 gcgcgtaata cgactcacta tagggcgaat tggagctcca ccgcggtg
//
