LOCUS       sequence-407424-        6813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Tick-borne powassan virus PrME protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6813)
  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 6813)
  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_POWV_PrME
FEATURES             Location/Qualifiers
     source          1..6813
                     /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(3518..3537)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3583..3638
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3637..3656)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3996..4012)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3996..4012)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4009..4031)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4020..4036
                     /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(4044..4050,4051..4068,4069..4074))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4089..4110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4227..4244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4398..4986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4478..4497)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5157..5948,5949..6017))
                     /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     5780..5799
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6018..6122)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6148..6603)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6285..6306)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6497..6516
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6730..6752
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6744..6761
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6745..6761
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6771..6790
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6771..6789
                     /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 tccggagtcg
     1441 attggacatg gatttttctg accatggccc tcatgaccat ggctatggcc accaccatcc
     1501 atagagatag ggagggctac atggtcatga gggcttccgg aagggacgcc gctagccaag
     1561 tgagggtgca aaacggcaca tgtgtgattc tggccaccga tatgggcgag tggtgtgagg
     1621 attccatcac ctacagctgc gtgaccattg accaagagga ggagcccgtc gacgtggact
     1681 gtttttgtag aggagtcgat agggtgaaac tcgagtatgg aagatgcggc agacaagccg
     1741 gaagcagagg caagagatcc gtcgtgatcc ccacccatgc ccaaaaagac atggtcggaa
     1801 ggggccatgc ttggctcaag ggcgacaaca ttagagacca cgtcacaaga gtggagggct
     1861 ggatgtggaa gaacaaactg ctgacagctg ccatcgtggc tctggcttgg ctcatggtgg
     1921 acagctggat ggctagagtg acagtgattc tgctggctct gtctctggga cccgtgtacg
     1981 ctacaaggtg cacccatctg gaaaatagag atttcgtgac cggcacacaa ggcacaacaa
     2041 gagtgtctct ggtgctcgag ctgggcggat gcgtgacaat cacagctgag ggcaagccta
     2101 gcatcgacgt gtggctcgag gacatcttcc aagagagccc cgccgagaca agggaatact
     2161 gcctccacgc taagctgaca aacaccaagg tggaggccag atgccctaca accggacccg
     2221 ccacactgcc cgaagaacac caagccaaca tggtctgcaa gagagaccag agcgataggg
     2281 gatggggcaa ccattgtggc tttttcggca aaggctccat cgtggcttgt gccaaattcg
     2341 agtgcgaaga ggccaagaag gccgtcggcc atgtgtacga cagcacaaaa atcacctacg
     2401 tggtgaaagt ggagccccac accggagact atctcgccgc taacgagacc aattccaata
     2461 gaaagagcgc ccagttcaca gtcgcctccg agaaggtgat tctgaggctg ggcgattatg
     2521 gcgacgtgtc tctgacatgc aaagtcgcta gcggcatcga tgtggctcag accgtggtga
     2581 tgtctctgga tagctccaag gatcatctgc ctagcgcttg gcaagtccac agagactggt
     2641 tcgaggatct ggctctgcct tggaagcaca aggacaacca agactggaac tccgtggaga
     2701 agctcgtgga gttcggcccc cctcacgccg tgaagatgga cgtctttaat ctgggagacc
     2761 agaccgctgt gctgctgaaa tctctggccg gcgtgcctct ggcctccgtg gagggacaaa
     2821 agtatcatct gaagagcggc catgtgacat gcgacgtggg cctcgagaag ctcaagctca
     2881 aaggcaccac ctacagcatg tgcgataagg ccaagttcaa gtggaaaagg gtgcccgtgg
     2941 acagcggaca cgatacagtc gtgatggagg tctcctacac cggctccgat aaaccttgca
     3001 gaatccccgt gagagctgtg gcccatggcg tccccgctgt gaatgtggcc atgctcatca
     3061 cccctaaccc caccatcgaa accaatggcg gcggctttat cgaaatgcaa ctgccccccg
     3121 gcgacaatat catctacgtg ggcgatctgt cccagcagtg gttccagaag ggctccacca
     3181 tcggcagaat gttcgaaaag accagaagag gactcgagag actgtccgtg gtcggcgagc
     3241 acgcttggga ctttggcagc gtcggcggag tcctcagctc cgtgggcaag gctatccaca
     3301 ccgtgctggg aggagccttc aacacactgt tcggaggagt gggcttcatc cctaagatgc
     3361 tgctgggcgt ggctctggtc tggctcggac tgaacgccag aaaccccacc atgtccatga
     3421 cctttctcgc cgtgggcgct ctgacactca tgatgaccat gggcgtcggc gcttgatgtc
     3481 ttcctggagc tccagtcatt gactataacg tctcagatct ttttccctct gccaaaaatt
     3541 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     3601 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     3661 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     3721 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     3781 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     3841 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     3901 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     3961 gatccctcga cggatcggcc gcaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     4021 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4081 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4141 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4201 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4261 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4321 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4381 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4441 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4501 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4561 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4621 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4681 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4741 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4801 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4861 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4921 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4981 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5041 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5101 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5161 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5221 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5281 gctgcaatga taccgcgaga cccacgctca ccggcaccag atttatcagc aataaaccag
     5341 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5401 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5461 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5521 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5581 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5641 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5701 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5761 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5821 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5881 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5941 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6001 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6061 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6121 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     6181 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     6241 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     6301 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     6361 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     6421 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     6481 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     6541 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     6601 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     6661 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     6721 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
     6781 cactataggg cgaattggag ctccaccgcg gtg
//
