LOCUS       sequence-433912-        5820 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Zika virus stable E dimer with fusion loop mutation and increased 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5820)
  AUTHORS   Phan TTN, Hvasta MG, Kudlacek ST, Thiono DJ, Tripathy A, Nicely NI, 
            de Silva AM, Kuhlman B
  TITLE     A conserved set of mutations for stabilizing soluble envelope 
            protein dimers from dengue and Zika viruses to advance the 
            development of subunit vaccines.
  JOURNAL   J Biol Chem. 2022 Jul;298(7):102079. doi: 10.1016/j.jbc.2022.102079.
            Epub 2022 May 26.
  PUBMED    35643320
REFERENCE   2  (bases 1 to 5820)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem."; date: "2022-07"; volume: "298(7):102079. doi"; pages: " 
            10.1016/j.jbc.2022.102079. Epub 2022 May 26"
COMMENT     Sequence Label: pZVsE_SC12
FEATURES             Location/Qualifiers
     source          1..5820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             3050..3073
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(3175..3194)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3240..3295
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3294..3313)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3656..3672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3656..3672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3669..3691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3680..3696
                     /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(3704..3710,3711..3728,3729..3734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3749..3770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3899..3916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4070..4658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4150..4169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4829..5620,5621..5689))
                     /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     5452..5471
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5690..5794)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccrgggggg gggggggggg gggggggggg
      541 ggggggggsg rggsggrgrg gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gkgggggtgc cgggcggggc
     1261 ggggccgcck cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tgcggccgtc
     1741 tcaggccgag ttcggtaccg ccaccatgaa gtgggtaacc tttatttccc ttctttttct
     1801 ctttagctcg gcttattcca tccgctgcat tggtgtgtca aatcgggatt tcgtagaggg
     1861 aatgtctggg ggcacatggg ttgacattgt cctcgaacat ggcaagtgcg tcacagtgat
     1921 gatgcaggat aaaccgacgg tagacatcga gcttgttact accactgtgt ccaatatggc
     1981 ggaagtgaga agctattgtt atgaggcgag catcagtgat atggcgagtg attcaaggtg
     2041 tcccacccaa ggcgaggcgt acttggataa acagagtgat acacaatacg tgtgtaagcg
     2101 aacactcgtt gacaggggtt gggggaacgg gtgcgacctc ttcggtaagg gaagtttggt
     2161 gacatgtgcg aagtttgcct gctctaagaa aatgactggg aagtccatac aaccagaaaa
     2221 tcttgaatat cgaattatgt tgtccgttca cggtagccag cattctggaa tgattgtcaa
     2281 tgatactggg cacgaaactg atgagaatcg agcgaaagta gagatcacgc ccaactcacc
     2341 tcgcgcggaa gcgacgttgg gagggtttgg ttccctggga ctggattgcg agcctagaac
     2401 tggactcgac ttttccgacc tgtactattt gacaatgaat aataagcatt ggttggtcca
     2461 caaggagtgg ttccatgata ttccgctgcc ctggcatgcg ggcgcggaca ctggaactcc
     2521 gcattggaat aacaaagaag ccctcgttga gtttaaagat gcccacgcca agaggcaaac
     2581 agttgtcgta ttgggctccc aggaaggttg ggtgcaccgg gctttggccg gagccctcga
     2641 agcagaaatg gacggagcga aaggtcgcct ttggcccgga cacctcaagt gcaggctgaa
     2701 gatggacaaa ctccgactca aaggggtttc ctatagcctt tgcacggcag cctttacatt
     2761 cacaaaaatc cccgccgaaa cgttgcacgg aactgtgacc gtggaggtgc agtatgccgg
     2821 cacggacggc ccttgcaaag tcccggctca aatggctgta gatatgcaaa cattgacccc
     2881 ggtcggacgg ctcattactg ccaatcctgt tatcacagaa tccacagaaa actcaaagat
     2941 gatgcttgag ctcgaccctc ccttcggaga ctcatatatt gtgattgggg taggtgaaaa
     3001 gaaaattact catcattggc atcgctctgg tggcagcagc ggcggcagcc atcaccacca
     3061 tcatcaccat cattagtaat gactcgagcg ataattcact cctcaggtgc aggctgccta
     3121 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     3181 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3241 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3301 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3361 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     3421 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3481 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     3541 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     3601 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     3661 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3721 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3781 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatcgat ccgctgcatt
     3841 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3901 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3961 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4021 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4081 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4141 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4201 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4261 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4321 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4381 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4441 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4501 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4561 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4621 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4681 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4741 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4801 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4861 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4921 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4981 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5041 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5101 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5161 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5221 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5281 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5341 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5401 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5461 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5521 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5581 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5641 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5701 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5761 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
