LOCUS       sequence-407382-        9087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of La Crosse virus GP protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9087)
  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 9087)
  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_LACV_GP
FEATURES             Location/Qualifiers
     source          1..9087
                     /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"
     primer_bind     complement(5792..5811)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5857..5912
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5911..5930)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6270..6286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6270..6286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6283..6305)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6294..6310
                     /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(6318..6324,6325..6342,6343..6348))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6363..6384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6501..6518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6672..7260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6752..6771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7431..8222,8223..8291))
                     /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     8054..8073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8292..8396)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8422..8877)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8559..8580)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8771..8790
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9004..9026
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9018..9035
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9019..9035
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9045..9064
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9045..9063
                     /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 caccatgatc tgcattctgg
     1441 tgctgatcac cgtggccgct gctagccccg tgtatcagag gtgtttccaa gacggcgcca
     1501 tcgtgaagca aaatccttcc aaggaagccg tgaccgaggt gtgtctgaag gacgatgtga
     1561 gcatgatcaa gaccgaagcc agatacgtga ggaatgctac cggcgtgttc tccaacaacg
     1621 tcgccattag gaagtggctg gtctccgact ggcacgattg cagacccaag aagatcgtcg
     1681 gcggacacat caacgtgatc gaggtgggcg acgacctctc cctccatacc gaaagctatg
     1741 tctgctccgc cgactgtacc attggcgtcg acaaggaaac cgcccaagtg agactgcaga
     1801 cagataccac aaaccatttc gagatcgctg gcaccaccgt gaaaagcgga tggttcaagt
     1861 ccaccaccta cattacactg gatcagacat gcgagcatct gaaggtgagc tgcggcccta
     1921 agagcgtgca gttccacgct tgcttcaatc agcacatgag ctgcgtgaga tttctgcata
     1981 gaaccattct gcccggaagc atcgccaact ccatctgtca aaatatcgaa attatcattc
     2041 tggtgaccct cacactgctc attttcattc tgctcagcat tctctccaag acatatatct
     2101 gttatctgct catgcctatt ttcattccca tcgcctacat ctacggcatc atttataaca
     2161 agagctgcaa aaagtgcaag ctgtgcggac tcgtgtacca tccctttaca gagtgcggca
     2221 cacattgtgt ctgcggcgct agatacgaca catccgatag aatgaagctc catagagcca
     2281 gcggactgtg tcccggctat aaatccctca gagccgctag agtcatgtgc aagagcaaag
     2341 gccccgccag cattctgagc atcatcaccg ccgtgctggt gctgaccttc gtcaccccca
     2401 tcaactccat ggtgctcgga gagtccaagg agaccttcga gctcgaggat ctgcccgacg
     2461 acatgctcga gatggcttct agaatcaata gctactacct cacatgcatt ctcaactacg
     2521 ccgtgagctg gggcctcgtc atcattggac tgctgatcgg actgctcttt aagaagtacc
     2581 agcatagatt cctcaacgtc tatgctatgt actgcgagga gtgcgacatg taccatgaca
     2641 aaagcggact gaagagacac ggcgacttca ccaacaagtg taggcagtgt acatgtggcc
     2701 agtatgagga cgccgccgga ctgatggctc atagaaagac ctacaactgt ctggtccagt
     2761 acaaagctaa gtggatgatg aatttcctca ttatttacat ctttctgatt ctgatcaagg
     2821 attccgctat cgtcgtgcaa gccgctggaa cagactttac cacatgcctc gagacagagt
     2881 ccatcaactg gaactgcacc ggcccctttc tcaacctcgg caattgtcag aagcagcaga
     2941 agaaggagcc ctataccaac atcgccacac agctcaaggg actgaaagct atctccgtgc
     3001 tggacgtgcc catcatcacc ggaatccccg atgacatcgc cggcgctctg agatacatcg
     3061 aagagaagga ggactttcac gtccagctca ccattgagta cgctatgctc tccaaatatt
     3121 gcgactacta cacccaattc agcgacaaca gcggatacag ccagaccaca tggagagtgt
     3181 acctcagaag ccacgatttc gaagcttgca tcctctaccc caaccaacac ttctgtaggt
     3241 gtgtcaagaa tggcgagaag tgctctagca gcaattggga tttcgccaac gagatgaagg
     3301 attattattc cggcaagcaa acaaagtttg ataaggatct gaatctcgcc ctcacagccc
     3361 tccatcacgc ttttagaggc acaagcagcg cctatattgc taccatgctc agcaagaagt
     3421 ccaatgacga cctcatcgcc tacaccaata agatcaaaac caagtttccc ggcaacgctc
     3481 tgctgaaagc catcattgac tatattgcct acatgaaaag cctccccggc atggccaatt
     3541 ttaaatacga cgagttctgg gacgagctcc tctacaagcc caatcccgct aaggcctcca
     3601 acctcgccag aggaaaggaa tcctcctata atttcaagct cgctatcagc agcaagtcca
     3661 tcaagacatg caagaatgtc aaagatgtgg cttgtctgag ccctaggtcc ggcgccattt
     3721 acgcttccat tatcgcttgc ggcgagccta acggaccctc cgtctataga aagccttccg
     3781 gcggcgtgtt tcaatccagc acagatagat ccatttattg tctgctcgac tcccactgtc
     3841 tcgaggagtt tgaagccatt ggacaagagg agctggacgc tgtgaagaag agcaaatgct
     3901 gggagatcga gtaccccgac gtcaagctga ttcaagaggg cgacggaaca aagagctgca
     3961 gaatgaagga ttccggcaac tgcaacgtcg ccaccaatag gtggcccgtg atccaatgtg
     4021 agaacgacaa gttttactac agcgagctgc agaaggacta cgacaaggcc caagacattg
     4081 gacattattg tctgagcccc ggctgcacaa cagtcagata tcccattaac cctaagcaca
     4141 tttccaattg taattggcaa gtgtctagaa gcagcattgc caagatcgac gtccacaaca
     4201 ttgaggatat tgaacagtat aagaaggcta ttacccaaaa gctgcagaca agcctctctc
     4261 tgtttaaata cgccaagaca aagaatctcc cccacatcaa acccatttat aagtatatca
     4321 ccattgaggg aacagagacc gctgagggca ttgagagcgc ctacattgag tccgaagtgc
     4381 ccgccctcgc cggcaccagc attggcttca agattaactc caaagaggga aaacatctgc
     4441 tggatgtgat tgcctatgtg aaatccgcct cctacagcag cgtgtacacc aaactctata
     4501 gcaccggacc cacatccggc atcaatacca agcacgacga actgtgtacc ggcccttgcc
     4561 ccgccaacat taaccaccaa gtgggctggc tgacattcgc tagggaaaga acaagcagct
     4621 ggggatgcga agagttcggc tgtctggctg tcagcgatgg ctgcgtcttt ggaagctgcc
     4681 aagacatcat caaagaagag ctgtccgtgt acagaaagga gacagaggag gtcacagacg
     4741 tcgagctctg tctgacattt agcgataaga cctattgcac caacctcaat cccgtgaccc
     4801 ctatcattac cgacctcttc gaagtgcagt ttaagacagt ggaaacatat tctctgccta
     4861 gaatcgtggc cgtgcaaaac catgagatca aaattggcca gatcaatgat ctcggcgtgt
     4921 atagcaaagg ctgtggcaac gtccagaagg tcaatggaac catctatggc aacggagtcc
     4981 ctagattcga ctatctgtgt catctggcta gcagaaagga agtgatcgtg aggaagtgct
     5041 ttgacaacga ttatcaagct tgcaagtttc tccaatcccc cgctagctat agactcgagg
     5101 aggatagcgg aacagtcacc atcatcgact ataagaagat tctcggcacc attaagatga
     5161 aggccattct gggcgacgtg aagtataaga cctttgccga ctccgtcgac atcaccgctg
     5221 aaggcagctg caccggctgt attaattgtt tcgagaatat ccactgtgag ctcacactgc
     5281 acacaacaat tgaagccagc tgccccatca agtccagctg caccgtgttc cacgatagaa
     5341 tcctcgtcac acccaacgag cacaaatacg ctctgaagat ggtgtgcacc gaaaagcccg
     5401 gcaacacact gacaattaaa gtctgtaata caaaggtcga ggctagcatg gctctggtgg
     5461 acgctaagcc tatcatcgag ctggctcccg tcgaccagac agcttacatt agagagaaag
     5521 acgagaggtg caagacatgg atgtgcagag tgagggacga aggactgcaa gtcatcctcg
     5581 agcccttcaa aaatctgttc ggaagctaca tcggcatttt ttataccttt attatctcca
     5641 tcgtggtgct gctggtcatc atctatgtgc tgctgcccat ttgcttcaag ctgagagaca
     5701 cactgagaaa gcatgaggac gcttacaaaa gagaaatgaa aattaggtga tgtcttcctg
     5761 gagctccagt cattgactat aacgtctcag atctttttcc ctctgccaaa aattatgggg
     5821 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     5881 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     5941 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     6001 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     6061 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     6121 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     6181 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     6241 tcgacggatc ggccgcaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     6301 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6361 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6481 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6541 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6601 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7081 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7381 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7561 atgataccgc gagacccacg ctcaccggca ccagatttat cagcaataaa ccagccagcc
     7621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8281 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8341 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8401 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     8461 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     8521 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     8581 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     8641 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     8701 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     8761 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     8821 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     8881 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     8941 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     9001 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     9061 agggcgaatt ggagctccac cgcggtg
//
