LOCUS       sequence-407396-        6816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of West Nile Virus PrME protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  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 6816)
  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_WNV_PrME
FEATURES             Location/Qualifiers
     source          1..6816
                     /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(3521..3540)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3586..3641
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3640..3659)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3999..4015)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3999..4015)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4012..4034)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4023..4039
                     /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(4047..4053,4054..4071,4072..4077))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4092..4113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4230..4247)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4401..4989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4481..4500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5160..5951,5952..6020))
                     /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"
     promoter        complement(6021..6125)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6151..6606)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6288..6309)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6500..6519
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6733..6755
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6747..6764
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6748..6764
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6774..6793
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6774..6792
                     /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 ggaggcaccg
     1441 ccggattcac cattctgctg ggactgatcg cttgcgccgg agccgtgacc ctcagcaact
     1501 tccaaggcaa ggtcatgatg accgtgaacg ccaccgacgt gaccgacgtc atcacaattc
     1561 ccaccgctgc cggcaagaat ctgtgcatcg tgagagccat ggatgtcggc tacctctgcg
     1621 aagataccat cacctacgaa tgccccgtgc tggctgccgg caatgatccc gaggacatcg
     1681 actgctggtg caccaagagc tccgtctacg tgaggtacgg aagatgcaca aaaacaagac
     1741 atagcagaag gagcagaagg agcctcacag tgcagacaca cggagagtcc accctcgcta
     1801 ataagaaggg agcttggctc gattccacca aagccaccag atatctggtc aagacagaga
     1861 gctggattct gaggaatccc ggctatgctc tcgtggctgc cgtcattggc tggatgctgg
     1921 gcagcaacac catgcaaagg gtcgtgttcg ccattctgct gctgctggtc gcccccgcct
     1981 actcctttaa ttgtctggga atgagcaata gagatttcct cgagggagtg agcggagcta
     2041 catgggtgga cctcgtgctc gagggagata gctgcgtcac cattatgtcc aaggacaaac
     2101 ccaccatcga cgtgaagatg atgaatatgg aggccgccaa tctggccgat gtgagatcct
     2161 actgttatct ggctagcgtg agcgatctga gcacaagggc tgcttgcccc acaatgggcg
     2221 aagcccacaa tgagaagagg gccgatcccg ccttcgtgtg caagcaaggc gtggtcgata
     2281 ggggctgggg caatggatgc ggactgtttg gcaaaggctc cattgacaca tgcgccaagt
     2341 tcgcttgcac caccaaggcc accggctgga tcatccagaa agaaaacatc aaatacgagg
     2401 tggccatttt cgtgcacggc cctacaaccg tggagtccca cggcaaaatt ggcgccaccc
     2461 aagccggcag attctccatt accccttccg ccccttccta tacactgaaa ctcggcgagt
     2521 acggcgaggt gacagtggat tgcgagccca gaagcggcat tgacacctcc gcctactatg
     2581 tcatgagcgt gggcgagaag tccttcctcg tccacagaga atggttcatg gatctgaatc
     2641 tgccttggag ctccgccgga tccacaacat ggagaaacag agagacactg atggagttcg
     2701 aggagcccca cgccacaaag cagagcgtcg tcgctctggg cagccaagag ggagctctcc
     2761 atcaagccct cgccggcgct attcccgtcg agttctcctc caacaccgtc aagctcacct
     2821 ccggccatct gaagtgtaga gtcaagatgg agaagctgca gctgaaaggc accacatatg
     2881 gagtctgcag caaggccttc aaattcgcca gaacacccgc tgataccggc catggaaccg
     2941 tggtcctcga actgcagtac accggcaccg atggcccttg caaggtccct atcagctccg
     3001 tggcttctct gaatgatctg acacccgtgg gcagactggt gacagtgaac ccctttgtga
     3061 gcgtggccac cgccaacagc aaagtgctga tcgaactcga gcctcctttc ggcgactcct
     3121 atatcgtcgt gggcagagga gaacagcaga tcaaccacca ctggcacaaa tccggctcct
     3181 ccattggcaa agcctttaca accacactga gaggcgccca aaggctcgct gccctcggag
     3241 ataccgcttg ggatttcgga agcgtgggag gcgtgtttac aagcgtgggc aaggctattc
     3301 accaagtgtt cggaggcgct tttaggtctc tgttcggcgg catgagctgg atcacccaag
     3361 gactgctggg cgctctgctg ctgtggatgg gcatcaacgc cagagacaga agcatcgcca
     3421 tgacctttct cgccgtcggc ggcgtgctgc tgtttctcag cgtcaacgtg catgcttgat
     3481 gtcttcctgg agctccagtc attgactata acgtctcaga tctttttccc tctgccaaaa
     3541 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     3601 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     3661 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     3721 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     3781 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     3841 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     3901 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     3961 ggagatccct cgacggatcg gccgcaattc gtaatcatgt catagctgtt tcctgtgtga
     4021 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4081 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4141 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4201 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4261 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4321 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4381 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4441 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4501 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4561 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4621 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4681 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4741 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4801 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4861 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4921 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4981 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5041 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5101 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5161 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5221 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5281 agtgctgcaa tgataccgcg agacccacgc tcaccggcac cagatttatc agcaataaac
     5341 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5401 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5461 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5521 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5581 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5641 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5701 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5761 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5821 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5881 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5941 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6001 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6061 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6121 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     6181 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     6241 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     6301 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     6361 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     6421 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     6481 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     6541 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     6601 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6661 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     6721 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
     6781 actcactata gggcgaattg gagctccacc gcggtg
//
