LOCUS       sequence-407399-        6810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Tick-borne encephalitis virus PrME protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6810)
  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 6810)
  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_TBEV_PrME
FEATURES             Location/Qualifiers
     source          1..6810
                     /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(3515..3534)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3580..3635
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3634..3653)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3993..4009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3993..4009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4006..4028)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4017..4033
                     /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(4041..4047,4048..4065,4066..4071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4086..4107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4224..4241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4395..4983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4475..4494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5154..5945,5946..6014))
                     /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     5777..5796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6015..6119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6145..6600)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6282..6303)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6494..6513
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6727..6749
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6741..6758
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6742..6758
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6768..6787
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6768..6786
                     /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 agcgccacag
     1441 actggatgag ctggctgctc gtgattacac tgctgggcat gacactggcc gctaccgtga
     1501 gaaaggagag ggatggaagc accgtcatta gagccgaagg aaaggatgct gccacccaag
     1561 tgagagtcga gaacggaaca tgcgtgattc tggctaccga catgggcagc tggtgcgacg
     1621 actctctgtc ctacgagtgc gtgaccatcg accaaggcga agagcccgtg gacgtggatt
     1681 gcttctgtag aaatgtcgac ggcgtctatc tggaatacgg cagatgcggc aagcaagagg
     1741 gctctagaac aagaagatcc gtgctgatcc ccagccatgc ccaaggcgag ctgaccggca
     1801 gaggacacaa gtggctcgag ggagattctc tgagaaccca tctgacaaga gtggaaggct
     1861 gggtgtggaa gaacaagctg ctggctctgg ccatggtcac agtcgtgtgg ctgaccctcg
     1921 agtccgtcgt cacaagagtg gctgtgctcg tcgtgctgct gtgcctcgct cccgtgtacg
     1981 cctctagatg cacacacctc gaaaatagag actttgtgac cggcacccaa ggcacaacca
     2041 gagtgaccct cgtgctggaa ctgggaggat gcgtgaccat tacagccgag ggcaagccca
     2101 gcatggatgt gtggctggac gccatttatc aagagaaccc cgccaagacc agagagtatt
     2161 gtctgcacgc caaactgtcc gacaccaaag tcgccgccag atgtcctacc atgggccccg
     2221 ccaccctcgc tgaagagcat caaggcggca ccgtgtgtaa aagggatcag agcgataggg
     2281 gatggggcaa ccattgcggc ctcttcggaa agggaagcat tgtggcttgc gtcaaggccg
     2341 cttgcgaagc caaaaagaag gccaccggcc acgtgtacga tgccaacaag atcgtctaca
     2401 ccgtcaaagt ggagccccat accggcgatt acgtggctgc caacgagacc cactccggca
     2461 gaaaaaccgc cagcttcaca atctcctccg aaaagaccat tctgaccatg ggcgaatacg
     2521 gcgacgtgtc tctgctctgc agagtggctt ccggcgtgga tctggcccaa accgtgatcc
     2581 tcgagctgga caagacagtc gaacatctgc ccacagcttg gcaagtgcat agagactggt
     2641 ttaacgacct cgctctgcct tggaaacatg agggcgccca gaactggaac aacgccgaga
     2701 gactcgtgga atttggcgct cctcatgccg tcaagatgga cgtgtacaat ctgggagacc
     2761 agaccggcgt gctgctgaag gctctggccg gcgtgcccgt ggctcacatc gagggcacca
     2821 agtaccatct gaagtccggc cacgtcacat gtgaggtcgg cctcgagaag ctgaagatga
     2881 agggactcac ctacacaatg tgcgacaaga ccaagttcac atggaagagg gcccctacag
     2941 atagcggaca tgacaccgtg gtgatggaag tgaccttttc cggcaccaag ccttgtagga
     3001 tccccgtgag ggctgtggcc catggcagcc ccgacgtgaa tgtcgccatg ctgatcaccc
     3061 ctaaccccac cattgagaac aacggcggag gcttcatcga gatgcagctg cctcccggcg
     3121 acaacatcat ttacgtcggc gagctgagcc accagtggtt tcaaaagggc agctccattg
     3181 gaagggtgtt ccaaaagacc aaaaagggaa tcgagaggct caccgtcatc ggagagcacg
     3241 cttgggactt cggatccgct ggcggctttc tgagcagcat tggcaaagcc gtccatacag
     3301 tgctgggcgg agccttcaac agcattttcg gcggagtggg cttcctccct aaactgctgc
     3361 tgggagtcgc tctggcttgg ctcggactca acatgagaaa ccccacaatg tccatgtcct
     3421 ttctgctggc tggcggactc gtgctcgcta tgaccctcgg cgtcggagcc tgatgtcttc
     3481 ctggagctcc agtcattgac tataacgtct cagatctttt tccctctgcc aaaaattatg
     3541 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     3601 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     3661 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     3721 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     3781 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     3841 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     3901 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     3961 ccctcgacgg atcggccgca attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     4021 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4081 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4141 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4201 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4261 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4321 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4381 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4441 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4501 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4561 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4621 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4681 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4741 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4801 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4861 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4921 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4981 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5041 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5101 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5161 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5221 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5281 gcaatgatac cgcgagaccc acgctcaccg gcaccagatt tatcagcaat aaaccagcca
     5341 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5401 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5461 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5521 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5581 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5641 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5701 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5761 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5821 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5881 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5941 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6001 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6061 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6121 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     6181 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6241 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6301 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6361 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6421 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6481 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6541 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6601 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     6661 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     6721 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     6781 tatagggcga attggagctc caccgcggtg
//
