LOCUS       sequence-407374-        6834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Murray valley encephalitis virus PrME 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6834)
  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 6834)
  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_MVEV_PrME
FEATURES             Location/Qualifiers
     source          1..6834
                     /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(3539..3558)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3604..3659
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3658..3677)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4017..4033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4017..4033)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4030..4052)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4041..4057
                     /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(4065..4071,4072..4089,4090..4095))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4110..4131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4248..4265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4419..5007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4499..4518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5178..5969,5970..6038))
                     /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(6039..6143)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6169..6624)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6306..6327)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6518..6537
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6751..6773
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6765..6782
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6766..6782
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6792..6811
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6792..6810
                     /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 ggcggatccg
     1441 agacctccgt gctgatgctg atcttcatgc tcatcggctt cgctgccgct ctgaagctgt
     1501 ccaccttcca aggcaagatc atgatgaccg tcaacgctac cgatatcgct gacgtcatcg
     1561 ccatccccac ccccaaggga cccaaccagt gctggattag ggccatcgat atcggcttca
     1621 tgtgcgacga caccatcacc tatgagtgtc ccaagctcga gagcggcaat gaccccgagg
     1681 acatcgattg ctggtgcgat aagcaagccg tctacgtcaa ctacggaaga tgtacaagag
     1741 ctagacactc caagaggtct agaagatcca tcaccgtcca aacacatgga gagtccaccc
     1801 tcgtgaacaa gaaggacgct tggctggatt ccaccaaggc tacaagatat ctgacaaaga
     1861 ccgagaattg gatcattaga aatcccggct acgctctggt ggctgtggtg ctgggatgga
     1921 tgctgggatc caataccggc cagaaggtga tcttcaccgt gctgctgctg ctggtggccc
     1981 ccgcttacag cttcaactgc ctcggaatga gcagcagaga cttcattgag ggcgcttccg
     2041 gcgctacatg ggtcgatctg gtgctcgagg gagacagctg tattaccatc atggctgccg
     2101 acaagcccac cctcgacatt agaatgatga acatcgaagc cacaaatctg gctctggtca
     2161 gaaactactg ctacgctgct accgtgagcg acgtgagcac cgtgagcaac tgtcctacaa
     2221 ccggcgaatc ccacaacacc aagagagccg accacaacta tctgtgcaag agaggcgtca
     2281 ccgacagagg ctggggcaac ggatgcggac tcttcggcaa gggctccatt gacacatgcg
     2341 ccaagttcac atgctccaat tccgccgccg gcagactgat tctccccgag gatatcaagt
     2401 acgaagtcgg cgtgttcgtg catggatcca ccgactccac aagccacggc aactactcca
     2461 cccagatcgg cgccaaccaa gccgtgaggt ttacaattag ccccaacgct cccgctatta
     2521 ccgctaagat gggcgactat ggcgaggtca ccgtggaatg cgagcctagg agcggactga
     2581 ataccgaggc ttactacgtc atgaccatcg gaaccaagca ctttctcgtg catagagagt
     2641 ggttcaatga tctcctcctc ccttggacaa gccccgcttc caccgagtgg aggaatagag
     2701 agattctggt ggagtttgag gagccccacg ccacaaagca gagcgtggtg gctctgggaa
     2761 gccaagaggg cgctctgcac caagctctgg ccggagccat tcccgtcgag tttagcagca
     2821 gcaccctcaa gctgacatcc ggacacctca agtgcagagt caagatggaa aagctgaagc
     2881 tgaagggcac cacctacgga atgtgtaccg agaagttcac cttcagcaag aaccccgctg
     2941 acaccggaca tggaaccgtg gtgctggaac tgcaatacac cggctccgac ggaccttgca
     3001 agatccctat tagctccgtg gcctccctca acgatatgac ccccgtggga agaatggtga
     3061 cagctaaccc ctacgtggct tccagcaccg ccaacgccaa ggtgctggtg gagatcgaac
     3121 cccccttcgg cgacagctat atcgtggtgg gaagaggcga caagcagatc aaccaccatt
     3181 ggcacaagga gggctcctcc attggcaagg ccttctccac cacactcaag ggagcccaaa
     3241 ggctggccgc tctgggagac acagcttggg atttcggcag cgtgggcggc gtgttcaact
     3301 ccatcggcaa agctgtgcat caagtgttcg gaggcgcttt cagaacactg tttggcggca
     3361 tgagctggat tagccccgga ctgctgggag ctctgctgct gtggatggga gtcaatgcta
     3421 gagataagtc catcgctctg gctttcctcg ccaccggagg cgtcctcctc tttctcgcca
     3481 ccaatgtgca tgcttgatgt cttcctggag ctccagtcat tgactataac gtctcagatc
     3541 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     3601 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     3661 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     3721 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     3781 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     3841 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     3901 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     3961 ctgtccctct tctcttatgg agatccctcg acggatcggc cgcaattcgt aatcatgtca
     4021 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4081 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4141 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4201 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4261 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4321 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4381 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4441 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4501 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4561 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4621 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4681 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4741 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4801 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4861 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4921 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4981 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5041 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5101 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5161 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5221 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5281 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggcacca
     5341 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5401 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5461 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5521 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5581 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5641 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5701 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5761 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5821 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5881 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5941 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6001 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6061 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6121 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     6181 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6241 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6301 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6361 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6421 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6481 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6541 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6601 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     6661 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6721 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6781 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccaccgc ggtg
//
