LOCUS       sequence-407397-        5856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Hepatitis C virus E2 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5856)
  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 5856)
  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_HCV_E2
FEATURES             Location/Qualifiers
     source          1..5856
                     /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(2561..2580)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2626..2681
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2680..2699)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3039..3055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3039..3055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3052..3074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3063..3079
                     /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(3087..3093,3094..3111,3112..3117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3132..3153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3270..3287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3441..4029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3521..3540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4200..4991,4992..5060))
                     /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     4823..4842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5061..5165)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5191..5646)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5328..5349)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5540..5559
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5773..5795
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5787..5804
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5788..5804
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5814..5833
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5814..5832
                     /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 caccatggag acacacgtga
     1441 ccggcggcag cgccggaaga acaacagccg gactggtggg actcctcaca cccggcgcca
     1501 aacagaacat ccagctcatc aacaccaacg gaagctggca catcaactcc acagctctga
     1561 actgcaatga gtctctgaac accggctggc tggccggact gttctatcag cacaagttca
     1621 atagcagcgg ctgccccgag agactggcta gctgtagaag gctgaccgac tttgctcaag
     1681 gctggggccc tatcagctat gccaacggca gcggcctcga tgagagacct tattgctggc
     1741 attacccccc tagaccttgc ggcatcgtcc ccgccaagtc cgtctgcgga cccgtgtatt
     1801 gctttacccc cagccccgtg gtcgtgggaa caaccgatag atccggagct cctacctata
     1861 gctggggcgc caacgacacc gacgtgttcg tgctgaataa caccagaccc cccctcggca
     1921 attggttcgg ctgcacatgg atgaatagca ccggcttcac caaagtgtgt ggcgctcccc
     1981 cttgcgtgat cggaggagtg ggcaataata cactgctgtg ccctacagac tgcttcagaa
     2041 agcaccccga ggccacatac tctagatgtg gcagcggccc ttggattacc cctaggtgta
     2101 tggtggacta tccctataga ctgtggcact atccttgcac catcaattac acaattttca
     2161 aggtgagaat gtacgtgggc ggagtggagc ataggctcga agccgcttgc aactggacaa
     2221 gaggcgagag atgcgacctc gaggatagag acagaagcga gctgtcccct ctgctgctga
     2281 gcaccacaca gtggcaagtg ctgccttgca gcttcacaac actgcccgct ctgagcaccg
     2341 gcctcatcca cctccaccag aacatcgtgg acgtgcagta tctgtacgga gtgggcagca
     2401 gcatcgctag ctgggccatc aaatgggagt acgtggtgct gctgtttctg ctcctcgccg
     2461 acgctagagt gtgtagctgt ctgtggatga tgctgctgat ctcccaagcc gaagcttgat
     2521 gtcttcctgg agctccagtc attgactata acgtctcaga tctttttccc tctgccaaaa
     2581 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     2641 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     2701 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     2761 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     2821 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     2881 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     2941 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     3001 ggagatccct cgacggatcg gccgcaattc gtaatcatgt catagctgtt tcctgtgtga
     3061 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4321 agtgctgcaa tgataccgcg agacccacgc tcaccggcac cagatttatc agcaataaac
     4381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5161 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     5221 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     5281 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     5341 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5401 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     5461 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5521 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5581 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5641 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5701 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     5761 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
     5821 actcactata gggcgaattg gagctccacc gcggtg
//
