LOCUS       sequence-407423-        7713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Eastern equine encephalitis virus E1E2 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7713)
  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 7713)
  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_EEE_E1E2
FEATURES             Location/Qualifiers
     source          1..7713
                     /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(4418..4437)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4483..4538
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4537..4556)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4896..4912)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4896..4912)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4909..4931)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4920..4936
                     /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(4944..4950,4951..4968,4969..4974))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4989..5010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5127..5144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5298..5886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5378..5397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6057..6848,6849..6917))
                     /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     6680..6699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6918..7022)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7048..7503)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7185..7206)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7397..7416
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7630..7652
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7644..7661
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7645..7661
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7671..7690
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7671..7689
                     /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 tctctggcca
     1441 cagtgatgtg cgtgctggct aacatcacat ttccttgcga tcagccccct tgcatgcctt
     1501 gctgttacga gaagaatcct catgagacac tgaccatgct ggaacagaac tacgacagca
     1561 gagcctatga ccagctgctg gatgccgctg tcaagtgcaa tgctagaaga accagaaggg
     1621 atctggatac ccacttcacc cagtataaac tcgctagacc ttacattgcc gactgtccta
     1681 attgcggaca cagcagatgc gacagcccta ttgctatcga ggaagtcaga ggagatgctc
     1741 acgctggcgt gatcagaatc caaacctccg ccatgttcgg cctcaaaaca gatggcgtcg
     1801 atctggccta catgagcttt atgaacggaa agacccagaa gtccatcaaa atcgacaatc
     1861 tccacgtgag gaccagcgct ccttgcagcc tcgtgtccca ccacggctac tacatcctcg
     1921 cccaatgccc ccccggagat accgtcacag tgggctttca cgacggcccc aatagacaca
     1981 catgcaccgt ggctcacaaa gtggaattta ggcccgtggg aagggagaag tacagacatc
     2041 cccccgagca tggcgtggaa ctgccttgca atagatatac ccacaaaaga gccgaccaag
     2101 gccactacgt ggaaatgcac caacccggcc tcgtggctga tcactctctg ctcagcatcc
     2161 actccgctaa ggtcaagatc acagtgccta gcggcgccca agtgaagtac tattgcaagt
     2221 gtcccgatgt gagagaagga accacaagca gcgattacac caccacatgc accgacgtca
     2281 agcaatgtag agcctacctc atcgacaaca agaaatgggt gtacaactcc ggcagactgc
     2341 ctagaggcga aggcgacacc ttcaagggca aactgcacgt cccttttgtg cccgtgaagg
     2401 ccaagtgtat cgccacactg gctcccgagc ctctggtcga gcacaagcat agaacactga
     2461 tcctccatct ctaccccgac catcccacac tgctgacaac aaggtctctg ggcagcgacg
     2521 ccaaccccac aaggcagtgg atcgagaggc ctacaacagt caacttcacc gtgaccggag
     2581 agggcctcga gtacacatgg ggcaatcatc cccctaaaag agtgtgggcc caagagtccg
     2641 gcgagggaaa tcctcacgga tggccccacg aagtggtggt ctactactac aatagatacc
     2701 ctctcaccac catcatcgga ctgtgtacat gcgtcgccat cattatggtg agctgcgtca
     2761 caagcgtgtg gctgctctgt agaacaagaa atctgtgcat caccccttac aagctcgccc
     2821 ccaacgccca agtgcctatc ctcctcgctc tgctgtgctg cattaagcct accagagccg
     2881 acgacaccct ccaagtgctg aactatctgt ggaacaacaa tcagaatttt ttttggatgc
     2941 aaaccctcat tcctctcgcc gctctgattg tctgcatgag aatgctgagg tgcctctttt
     3001 gttgcggacc cgcctttctg ctggtctgcg gagctctcgg agctgctgcc tatgaacaca
     3061 ccgccgtcat gcccaacaaa gtgggaatcc cctacaaggc cctcgtggaa agacccggct
     3121 atgctcccgt gcatctgcag atccagctcg tgaatacaag aattatccct agcaccaacc
     3181 tcgagtacat cacatgcaag tataaaacaa aagtccctag ccccgtggtg aagtgttgcg
     3241 gagccaccca gtgtacaagc aagccccacc ccgattacca gtgccaagtg tttaccggcg
     3301 tctacccctt catgtgggga ggagcctact gcttctgcga cacagagaac acacagatgt
     3361 ccgaggccta tgtggagaga tccgaggagt gttccatcga tcacgccaag gcctacaaag
     3421 tccataccgg cacagtccaa gccatggtga acatcaccta cggaagcgtg tcttggagaa
     3481 gcgccgatgt ctacgtgaac ggcgaaaccc ccgccaagat tggcgacgcc aagctgatca
     3541 ttggacctct gagctccgct tggtccccct tcgacaacaa ggtggtggtg catggccacg
     3601 aggtctataa ctacgatttc cccgagtatg gcaccggcaa agccggctcc tttggagacc
     3661 tccaatctag aacctccaca agcaacgacc tctacgccaa cacaaatctg aaactgcaga
     3721 gacctcaagc cggcatcgtg catacaccct ttacccaagc cccttccggc ttcgagagat
     3781 ggaagaggga caaaggagct cctctcaacg atgtggcccc ctttggctgc tccattgctc
     3841 tcgagcctct gagagctgag aactgtgctg tgggctccat ccctatcagc attgacatcc
     3901 ccgacgctgc tttcacaagg attagcgaga cacccaccgt gagcgatctc gagtgcaaga
     3961 tcaccgagtg cacctatgcc tccgacttcg gaggaattgc caccgtcgcc tacaaatcct
     4021 ccaaggccgg caactgtcct atccatagcc cttccggcgt cgccgtgatc aaagagaacg
     4081 acgtgacact ggccgagtcc ggaagcttca ccttccactt cagcaccgcc aacattcacc
     4141 ccgcttttaa actgcaagtg tgcaccagcg ccgtgacatg caaaggcgac tgtaagcccc
     4201 ccaaggacca tatcgtggat taccccgccc agcacacaga gtcctttaca agcgccatca
     4261 gcgccacagc ttggagctgg ctcaaagtgc tggtgggcgg cacatccgcc ttcatcgtgc
     4321 tcggactcat tgccaccgcc gtggtggctc tggtgctgtt tttccacaga cattgatgtc
     4381 ttcctggagc tccagtcatt gactataacg tctcagatct ttttccctct gccaaaaatt
     4441 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     4501 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     4561 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     4621 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     4681 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     4741 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     4801 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     4861 gatccctcga cggatcggcc gcaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     4921 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4981 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5041 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5101 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5161 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5221 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5281 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5701 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5881 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6121 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6181 gctgcaatga taccgcgaga cccacgctca ccggcaccag atttatcagc aataaaccag
     6241 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6301 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6361 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6421 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6481 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6541 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6601 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6661 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6721 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6781 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6841 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6901 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6961 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7021 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     7081 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     7141 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     7201 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     7261 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     7321 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     7381 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     7441 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     7501 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7561 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     7621 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
     7681 cactataggg cgaattggag ctccaccgcg gtg
//
