LOCUS       sequence-268918-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon-optimized HSV-1 UL33 with N-terminal HAStrep tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  AUTHORS   Wyler E, Franke V, Menegatti J, Kocks C, Boltengagen A, Praktiknjo 
            S, Walch-Ruckheim B, Bosse J, Rajewsky N, Grasser F, Akalin A, 
            Landthaler M
  TITLE     Single-cell RNA-sequencing of herpes simplex virus 1-infected cells 
            connects NRF2 activation to an antiviral program.
  JOURNAL   Nat Commun. 2019 Oct 25;10(1):4878. doi: 10.1038/s41467-019-12894-z.
  PUBMED    31653857
REFERENCE   2  (bases 1 to 5631)
  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: "2019-10-25"; volume: "10(1):4878. doi"; pages: " 
            10.1038/s41467-019-12894-z"
COMMENT     Sequence Label: pcDNA5FRTTO-NHASt-codoptUL33
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     255..279
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             373..399
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             430..453
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             490..513
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(969..986)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    975..1199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1332..1351)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1483..1530
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             1538..2560
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             1538..2560
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    2690..2811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2727..2746)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2781..2800
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2860..2876)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2860..2876)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2873..2895)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2884..2900
                     /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(2908..2914,2915..2932,2933..2938))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2953..2974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3091..3108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3262..3850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3342..3361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4021..4812,4813..4881))
                     /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     4644..4663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4882..4986)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5061..5080)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5252..5631
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctccct atcagtgata gagatctccc tatcagtgat
      241 agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
      301 tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct agcgtttaaa
      361 cttaagctta tgtacccata cgatgttcct gactatgccg gaaccgagct gggttccacg
      421 atggcaagct ggagccaccc gcagttcgag aaaggtggag gttccggagg tggatcggga
      481 ggtggatcgt ggagccaccc gcagttcgaa aaagggaacc ttttagcaga ggtaccgatg
      541 gccggaagag agggcaggac aaggcagagg accctgaggg acacaatccc tgattgcgcc
      601 ctgagaagcc agaccctgga gagcctggac gccagatacg tgagcagaga cggcgcccac
      661 gatgccgccg tgtggttcga ggatatgaca cccgccgagc tggaggtggt gtttcctaca
      721 acagacgcca agctgaatta cctgtccagg acacagagac tggcctccct gctgacctac
      781 gccggcccaa tcaaggcccc tgacgacgcc gccgcccctc aaacacccga taccgcctgc
      841 gtgcacggcg agctgctggc tgctaagagg gagaggtttg ccgccgtgat caacaggttc
      901 ctggacctgc accagatcct gtgagcggcc gctcgagtct agagggcccg tttaaacccg
      961 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1021 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1081 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1141 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     1201 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1261 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1321 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1381 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1441 cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag
     1501 ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac tcaccgcgac
     1561 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     1621 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     1681 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     1741 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta
     1801 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     1861 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca
     1921 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     1981 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     2041 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     2101 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     2161 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     2221 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     2281 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat
     2341 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     2401 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     2461 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     2521 aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac gagatttcga
     2581 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     2641 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     2701 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2761 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2821 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2881 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2941 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3001 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3061 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3121 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3181 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3241 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3301 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3361 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3601 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3661 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3721 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3781 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4141 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4981 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     5041 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     5101 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     5161 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     5221 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     5281 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     5341 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     5401 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     5461 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     5521 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     5581 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat g
//
