LOCUS       sequence-345858-        4804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of KSHV Orf 50 RTA phosphomutant generated by 
            amino acid substitutions S47A, S65A, T75A introduced to WT RTA 
            sequence from Nakamura et al., JVI 2003. 77:4205-20 (PMID: 
            12634378)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4804)
  AUTHORS   Alzhanova D, Meyo JO, Juarez A, Dittmer DP
  TITLE     The ORF45 Protein of Kaposi Sarcoma-Associated Herpesvirus Is an 
            Inhibitor of p53 Signaling during Viral Reactivation.
  JOURNAL   J Virol. 2021 Nov 9;95(23):e0145921. doi: 10.1128/JVI.01459-21. Epub
            2021 Sep 15.
  PUBMED    34523970
REFERENCE   2  (bases 1 to 4804)
  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: "J Virol."; 
            date: "2021-11-9"; volume: "95(23):e0145921. doi"; pages: " 
            10.1128/JVI.01459-21. Epub 2021 Sep 15"
COMMENT     Sequence Label: pDD2601
FEATURES             Location/Qualifiers
     source          1..4804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     159..177
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(215..237)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     337..356
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     550..572
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     564..581
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     565..581
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      627..636
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2769..2785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2769..2785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2782..2804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2793..2809
                     /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(2817..2823,2824..2841,2842..2847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2862..2883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3000..3017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3171..3759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3251..3270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3930..4721,4722..4790))
                     /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     4553..4572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4791..4804,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
      181 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
      241 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
      301 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
      361 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      421 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      481 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      541 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtacct
      601 cgcgaatgca tctagatgaa ttcgccgcca ccatggcgca agatgacaag ggtaagaagc
      661 ttcggcggtc ctgtgtggaa agcttcgtcg gcctctcgga cgaactgaag gcccaactct
      721 accagtgtgt gctccttata aatgacgcat acgaaacaat ctacgatccc gctgacctaa
      781 atagagtggt ggaagatgtg tgcattcgga ttatgaaaga atgtgccaag cttggtgcgc
      841 tatgtggtct gtttgcagac attaacatgt ttaacctttt ctgctttttt cgtgcctctc
      901 gaatgaggac caaaggcgcg gccgggtaca acgtgccatg cgcagaggca tcccaaggca
      961 ttattcggat cctcacggag aggatcttat tctgcacaga aaaggcattt ctgacagccg
     1021 catgcagcgg ggtgagcctg cctccagcca tatgtaagct actacacgaa atatacactg
     1081 aaatgaaggc caaatgcctg ggggcctgga ggcgactcgt ctgcaatcgg aggcccatta
     1141 tgatattaac ctcttcccta ctgaagctct acaacacgta cgataccgcc gggctgctct
     1201 ctgagcagtc cagggccctc tgccttttgg ttttccaacc ggtctacctt ccgaggatta
     1261 tggcgccgct ggagatcatg accaagggtc agctcgcccc tgaaaacttt tacagcatca
     1321 ccggttctgc tgagaaacgc cggccaatta ccaccggcaa ggtcactgga ctgtcctatc
     1381 caggaagcgg tctcatgcca gaatctttaa ttttgccaat cctggagcca ggactgttgc
     1441 cggcttccat ggtagacctc agcgatgtgc tggcaaaacc cgccgttatt ctgagcgccc
     1501 ctgccctgag ccagtttgtc attagcaaac cccatcccaa catgccgcac accgtcagca
     1561 tcatcccctt taacccatcg ggtacagacc cggcgtttat tagtacgtgg caggccgcgt
     1621 cacagaatat ggtgtacaac acatccaccg cgcccttaaa accggccacc ggtagttcac
     1681 agacggtgtc agtcaaggcg gttgctcaag gggccgtgat tactgcgaca acggtgccgc
     1741 aggcaatgcc agcgcggggt accggagggg agttgcctgt aatgtcagcg tccactcctg
     1801 caagagatca ggtcgctgca tgttttgtcg cagagaacac cggagattct cccgacaacc
     1861 cgagctcttt cctgacgtca tgtcaccctt gcgatccgaa cacggttata gtggcccagc
     1921 aatttcaacc accgcaatgc gttacgttgt tgcaggttac ctgtgccccc tcttcgacac
     1981 caccccccga ttcaacagtc cgggccccgg tggtgcagtt gccaacagta gtccctctgc
     2041 cggccagcgc gttcctcccg gcgctcgccc aaccagaagc ctcgggcgaa gagcttccgg
     2101 gcggtcatga cggagaccaa ggtgtgccgt gtagagattc aacggcggcg gctacggcgg
     2161 cagaggcgac aacacccaaa cgaaagcaga gaagcaaaga gaggagctca aagaagcgta
     2221 aggctttgac cgtgccagaa gccgacacca cgccatcgac cacgacacct ggtacctctt
     2281 tgggatcaat taccaccccc caggatgtgc acgccacgga tgtcgccacg tctgagggac
     2341 catcggaggc acaacccccg ctactgtcgt tacccccgcc actggacgta gatcagagtc
     2401 tattcgccct gttagacgaa gcgggccctg aaacatggga tgtcgggtcg cctctctccc
     2461 ccactgacga cgcgctgttg tccagtattc tgcaaggact gtaccagctg gacacgccac
     2521 cgcctctgcg gtcaccctcc cccgcttcct tcggcccgga gtctccggcg gatataccgt
     2581 caccttctgg tggagagtat acgcaactgc aaccggtcag ggcgacctcg gcgacgcccg
     2641 ctaacgaggt acaggagtcc ggcacactgt accagctgca ccaatggcgt aattacttcc
     2701 gagactctag aatcggatcc cgggcccgtc gactgcagag gcctgcatgc aagcttggcg
     2761 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2821 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3001 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3061 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3601 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4321 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4381 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4441 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4501 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4561 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4621 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4681 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4741 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4801 tttc
//
