LOCUS       sequence-407372-        7728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Semliki Forest virus E1E2 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7728)
  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 7728)
  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_SFV_E1E2
FEATURES             Location/Qualifiers
     source          1..7728
                     /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"
     CDS             3055..3066
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4433..4452)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4498..4553
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4552..4571)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4911..4927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4911..4927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4924..4946)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4935..4951
                     /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(4959..4965,4966..4983,4984..4989))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5004..5025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5142..5159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5313..5901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5393..5412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6072..6863,6864..6932))
                     /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     6695..6714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6933..7037)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7063..7518)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7200..7221)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7412..7431
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7645..7667
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7659..7676
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7660..7676
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7686..7705
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7686..7704
                     /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 caccatgggc agcgcccctc
     1441 tgattaccgc tatgtgcgtc ctcgccaatg ccacattccc ttgctttcag cctccttgcg
     1501 tgccttgctg ttacgagaat aacgccgaag ctacactgag gatgctcgaa gataacgtcg
     1561 atagacccgg ctactacgat ctgctgcaag ccgctctgac atgtagaaac ggcaccagac
     1621 atagaaggag cgtcagccag cacttcaatg tctacaaagc tacaagacct tacatcgcct
     1681 actgcgccga ttgcggagcc ggacatagct gccatagccc cgtggccatt gaagctgtga
     1741 ggagcgaggc taccgacggc atgctgaaga tccaattctc cgcccaaatt ggaatcgaca
     1801 agagcgacaa ccacgactat accaagatta gatacgccga cggacacgcc atcgaaaatg
     1861 ccgtgagatc ctctctgaag gtcgccacat ccggcgactg cttcgtccac ggaaccatgg
     1921 gacactttat tctggctaaa tgcccccccg gcgagtttct ccaagtctcc atccaagaca
     1981 ccagaaacgc cgtgagagct tgcagaatcc aataccacca cgatccccag cccgtgggaa
     2041 gggagaagtt caccattaga ccccactacg gcaaagagat cccttgcaca acataccagc
     2101 agaccacagc cgagacagtg gaggagatcg acatgcatat gccccccgat acccccgaca
     2161 gaacactgct gagccaacaa agcggaaacg tgaagatcac cgtgggaggc aagaaggtga
     2221 agtacaactg cacatgcgga accggaaatg tcggcaccac aaacagcgac atgaccatta
     2281 acacatgcct catcgagcag tgtcacgtgt ccgtgaccga ccataagaaa tggcagttta
     2341 actccccttt cgtccccaga gccgacgagc ccgccagaaa aggaaaggtg cacatccctt
     2401 tccctctgga caacattaca tgcagagtcc ccatggccag agagcccacc gtcatccacg
     2461 gaaagaggga ggtgacactg cacctccacc ccgaccaccc tacactgttc agctatagaa
     2521 ccctcggcga ggatcctcag taccatgagg aatgggtgac agctgccgtc gagaggacca
     2581 ttcccgtgcc cgtggatgga atggagtacc actggggcaa caatgacccc gtgagactgt
     2641 ggagccaact gaccaccgaa ggaaaacccc acggctggcc ccaccaaatc gtgcaatact
     2701 attacggcct ctatcccgcc gccacagtct ccgctgtggt gggaatgtct ctgctggctc
     2761 tgatctccat cttcgccagc tgctatatgc tggtggccgc cagaagcaaa tgcctcaccc
     2821 cctatgctct gacccccggc gccgctgtcc cttggacact gggcattctg tgctgcgctc
     2881 ccagagccca cgctgctagc gtcgctgaga caatggccta tctctgggac cagaaccaag
     2941 ctctgttctg gctggaattt gctgctcccg tggcttgcat tctgatcatt acctactgtc
     3001 tgaggaacgt gctctgctgc tgtaagagcc tctcctttct ggtgctcctc tctctgggag
     3061 ctacagccag agcttacgag cactccaccg tgatgcctaa cgtcgtgggc tttccttaca
     3121 aagcccacat cgagagaccc ggctacagcc ctctgacact gcaaatgcaa gtggtcgaga
     3181 cctctctgga acccacactg aatctggaat acattacatg cgagtacaag accgtggtgc
     3241 ctagccccta cgtgaagtgc tgcggcgcct ccgagtgctc cacaaaagag aagcccgatt
     3301 accagtgcaa ggtctacacc ggcgtgtatc ctttcatgtg gggaggcgcc tactgctttt
     3361 gcgatagcga gaacacccag ctcagcgagg cttacgtgga cagaagcgat gtgtgcagac
     3421 atgatcacgc tagcgcctac aaggcccata cagcctctct gaaggccaaa gtgagggtca
     3481 tgtacggcaa cgtgaatcag accgtcgacg tgtacgtcaa cggagatcac gctgtgacca
     3541 tcggcggaac ccagttcatt tttggccctc tgagcagcgc ttggacccct ttcgacaata
     3601 aaatcgtggt ctacaaggac gaagtgttca accaagactt ccctccctat ggaagcggcc
     3661 agcccggcag attcggcgat atccagagca gaaccgtcga gtccaatgac ctctacgcta
     3721 acaccgctct gaagctggct aggccttccc ccggcatggt gcatgtgccc tacacccaga
     3781 caccctccgg attcaaatac tggctgaagg agaagggcac cgctctgaac acaaaagccc
     3841 ccttcggatg tcagatcaag accaaccccg tgagagccat gaattgcgcc gtgggaaaca
     3901 tccccgtgtc catgaacctc cccgacagcg ccttcacaag aatcgtcgaa gctcccacca
     3961 tcatcgacct cacatgcaca gtggctacat gcacccactc ctccgatttc ggcggcgtgc
     4021 tcacactcac atacaagacc aacaagaacg gcgactgttc cgtgcacagc cacagcaacg
     4081 tggccacact gcaagaggcc accgccaagg tcaagaccgc cggcaaagtg acactgcact
     4141 tctccacagc cagcgcctcc cctagctttg tggtgtctct gtgtagcgcc agagccacat
     4201 gcagcgccag ctgtgaacct cccaaagatc acatcgtccc ctacgctgct agccactcca
     4261 atgtcgtgtt ccccgacatg agcggaaccg ctctgagctg ggtgcagaaa atcagcggcg
     4321 gcctcggcgc ttttgctatc ggagccattc tcgtgctggt cgtggtcaca tgcatcggac
     4381 tgagaagatg atgtcttcct ggagctccag tcattgacta taacgtctca gatctttttc
     4441 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     4501 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     4561 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     4621 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     4681 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     4741 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     4801 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     4861 ctcttctctt atggagatcc ctcgacggat cggccgcaat tcgtaatcat gtcatagctg
     4921 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4981 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5041 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5101 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5161 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5221 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5281 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5341 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5401 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5461 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5521 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5581 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5641 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5701 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5761 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5821 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5881 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5941 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6001 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6061 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6181 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc accagattta
     6241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6361 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6661 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6721 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6841 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6901 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6961 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7021 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     7081 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     7141 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     7201 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     7261 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     7321 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     7381 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     7441 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     7501 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     7561 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     7621 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     7681 gcgcgtaata cgactcacta tagggcgaat tggagctcca ccgcggtg
//
