LOCUS       sequence-407418-        6837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Japanese encephalitis virus PrME protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6837)
  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 6837)
  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_JEV_prME
FEATURES             Location/Qualifiers
     source          1..6837
                     /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(3542..3561)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3607..3662
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3661..3680)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4020..4036)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4020..4036)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4033..4055)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4044..4060
                     /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(4068..4074,4075..4092,4093..4098))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4113..4134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4251..4268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4422..5010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4502..4521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5181..5972,5973..6041))
                     /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     5804..5823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6042..6146)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6172..6627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6309..6330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6521..6540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6754..6776
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6768..6785
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6769..6785
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6795..6814
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6795..6813
                     /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 ggaggcaacg
     1441 agggatccat catgtggctc gcctccctcg ccgtggtgat tgcctatgcc ggagccatga
     1501 agctgagcaa ctttcaaggc aagctgctga tgacaatcaa taacaccgat atcgccgatg
     1561 tcattgtgat ccccacctcc aaaggcgaga acagatgctg ggtgagagcc attgacgtcg
     1621 gctacatgtg cgaggacacc attacctacg agtgccctaa actgaccatg ggcaatgatc
     1681 ccgaggacgt cgactgttgg tgtgataacc aagaggtcta cgtccaatac ggaagatgta
     1741 ccagaacaag gcacagcaag aggagcagaa ggtccgtgag cgtgcaaacc catggagagt
     1801 cctctctggt caacaaaaag gaagcttggc tcgacagcac caaagccaca aggtatctga
     1861 tgaaaaccga gaactggatc attagaaacc ccggctacgc cttcctcgcc gctacactgg
     1921 gctggatgct gggctccaat aatggacaaa gggtggtgtt taccattctg ctgctgctgg
     1981 tggcccccgc ctatagcttt aattgtctgg gcatgggcaa tagggacttc attgagggcg
     2041 ctagcggcgc tacatgggtg gatctggtgc tggaaggcga tagctgcctc accattatgg
     2101 ccaatgacaa gcccacactg gacgtgagga tgatcaacat tgaggccagc cagctcgccg
     2161 aagtgagatc ctactgttac cacgccagcg tgacagatat cagcacagtg gccagatgtc
     2221 ctaccaccgg cgaggctcat aatgagaaga gggctgacag cagctacgtg tgtaaacaag
     2281 gcttcaccga cagaggctgg ggaaatggat gtggactgtt cggcaagggc tccatcgaca
     2341 catgcgccaa gttcagctgc acctccaagg ctatcggaag aaccattcag cccgagaaca
     2401 tcaagtacga agtcggcatt ttcgtccacg gaacaaccac ctccgaaaac cacggcaatt
     2461 actccgctca agtgggagcc tcccaagccg ccaaattcac aattacaccc aacgccccca
     2521 gcatcacact caagctgggc gattacggag aggtgaccct cgactgcgaa cctagatccg
     2581 gactgaacac cgaagccttt tacgtgatga ccgtgggaag caagtccttt ctggtgcata
     2641 gggaatggtt ccacgacctc gccctccctt ggacatcccc cagctccacc gcttggagaa
     2701 atagagaact gctgatggag ttcgaggagg ctcacgctac aaagcagagc gtcgtggccc
     2761 tcggctccca agaaggagga ctgcaccaag ccctcgccgg cgccattgtg gtcgaatatt
     2821 ccagctccgt gaagctcaca agcggacatc tgaagtgtag gctcaagatg gacaagctcg
     2881 ctctgaaagg aaccacatac ggaatgtgca ccgagaagtt cagcttcgct aagaaccccg
     2941 ccgataccgg ccacggcacc gtggtcatcg agctgagcta tagcggcagc gatggccctt
     3001 gcaaaatccc tattgtcagc gtggcctctc tgaacgacat gacccccgtg ggcagactcg
     3061 tcacagtgaa tcccttcgtc gctacctcct ccgccaatag caaggtgctc gtcgagatgg
     3121 aacctccttt tggcgactcc tacatcgtcg tcggcagagg cgataagcag atcaaccacc
     3181 actggcataa ggctggaagc accctcggca aagcttttag caccacactg aagggcgctc
     3241 aaagactggc tgccctcgga gatacagctt gggatttcgg ctccatcgga ggagtcttta
     3301 acagcatcgg caaggccgtg caccaagtgt ttggcggcgc tttcagaaca ctgtttggcg
     3361 gcatgagctg gatcacccaa ggactgatgg gcgctctgct gctctggatg ggagtgaacg
     3421 ccagagatag gagcatcgct ctggcctttc tcgctaccgg cggcgtgctg gtgttcctcg
     3481 ccacaaatgt ccacgcctga tgtcttcctg gagctccagt cattgactat aacgtctcag
     3541 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     3601 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     3661 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     3721 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     3781 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     3841 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     3901 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     3961 tagctgtccc tcttctctta tggagatccc tcgacggatc ggccgcaatt cgtaatcatg
     4021 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4081 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4141 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4201 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4261 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4321 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4861 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4981 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5041 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5101 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5161 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5221 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5281 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggca
     5341 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5401 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5461 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5521 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5581 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5641 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5701 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5761 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5821 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5881 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5941 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6001 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6061 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6121 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     6181 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6241 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6301 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     6361 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     6421 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     6481 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     6541 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6601 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     6661 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6721 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6781 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctccac cgcggtg
//
