LOCUS       sequence-258856-        8901 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Firefly Luciferase reporter for K8.1 promoter from KSHV with the 
            lytic origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8901)
  AUTHORS   Nandakumar D, Glaunsinger B
  TITLE     An integrative approach identifies direct targets of the late viral 
            transcription complex and an expanded promoter recognition motif in 
            Kaposi's sarcoma-associated herpesvirus.
  JOURNAL   PLoS Pathog. 2019 May 16;15(5):e1007774. doi: 
            10.1371/journal.ppat.1007774. eCollection 2019 May.
  PUBMED    31095645
REFERENCE   2  (bases 1 to 8901)
  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: "PLoS 
            Pathog."; date: "2019-05-16"; volume: "15(5):e1007774. doi"; pages: 
            " 10.1371/journal.ppat.1007774. eCollection 2019 May"
COMMENT     Sequence Label: K8.1Pr-pGL4.16-OriLyt
FEATURES             Location/Qualifiers
     source          1..8901
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(1045..1836,1837..1905))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    2010..2058
                     /note="synthetic polyadenylation signal"
     misc_feature    2072..2163
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2112..2131
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2312..2321
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2318..3967
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             3974..4021
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             4025..4144
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    4193..4314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4204..4223)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4258..4277
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4508..4865
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4508..4699
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4531..4551)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4716..4851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4778..4797
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4896..5933
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5912..5920
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5957..6005
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6161..6178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6332..6920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6412..6431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccccccggag ggggatcccg gcgcgccacc ctcccccgga gggggatccc ggcgcgccac
       61 cctcccccgg agggggatcc cggcgcgcca ccctcccccg gagggggatc ccggcgcgcc
      121 accctccccc ggagggggat cccggcgcgc caccctcccc ggcaacaacc tgttgccatg
      181 tatggcgatt tgtatcagtc acaagcacac aacccctgct agtattaatg gtgtttaaaa
      241 cgttctacac gtacggcgga ccgcatccgt cgcaagcacg cgcatataac ccccaaatgc
      301 accatgatga gaagcacagc cacgcgtcaa aaaactttaa aaacatcgtt atccaatatc
      361 attaaaaacc acaccgaaat ttacacaggt agcacgtcac cgtgttagtg tcacccactg
      421 tacacaaggc gtgtcgtata tgtagtatag gtatttgatg aggcggaagc atatcccgct
      481 tccagcgaac ggaaataaga atcatccgtt ccagcattta ttcaaagggg gcacagagga
      541 ttcacattgt ttagagagag tttttcttag tcaccattcc atacttgggc agtattggcc
      601 tacgatttgg gcgacgtttc aggctggtct attctccgtc cacttttccc cggctattct
      661 gtcccagcat aggctcttga aataaacaat gtttaccgag taaaaggttc cactcaccct
      721 catttgtcgt tgcacccatc ccccctttgc ttaatcaccc gaaaactaga ggacacggat
      781 ggaaaacata tcgcacgcgg gttgtttgaa agtcaacagc tacttgtttt taatgaggac
      841 agatttgggc acaggccaga gggtaaagcc ctacgtgtgc gcgggggggg tgtatacgct
      901 gcgaaaacct gcacggtgca taacacccag ggcgtcacgt cacatatctc tgtgcaccca
      961 agtggttgtt caaccgttgt tttttggatg atttttccgc accggctttt ttgtgggcgc
     1021 gcataggtcg gtccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc
     1081 gatctgccta tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat
     1141 tcgtgagggc ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc
     1201 ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc
     1261 tgctactttg tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag
     1321 ttcgccagtg agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg
     1381 ctcgtcgttc ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg
     1441 atcacccata ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag
     1501 taagttggcc gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt
     1561 catgccatcc gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga
     1621 gtagtgtata cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc
     1681 acatagcagt actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc
     1741 aaggatcttg ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc
     1801 ttcagcatct tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc
     1861 cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca
     1921 atattattga agcatttatc agggttacta gtacgtctct caaggataag taagtaatat
     1981 taaggtacgg gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt
     2041 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
     2101 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     2161 tctggcctaa ctggccggta cctgtgccgt cgtccgggag aaccatgcca gactttgtgt
     2221 ggtaagaagg aattgttatc cggcagcaat attaaaggga ccgaagttaa tcccttaatc
     2281 ctaagcttgg caatccggta ctgttggtaa agccaccatg gaagatgcca aaaacattaa
     2341 gaagggccca gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa
     2401 agccatgaag cgctacgccc tggtgcccgg caccatcgcc tttaccgacg cacatatcga
     2461 ggtggacatt acctacgccg agtacttcga gatgagcgtt cggctggcag aagctatgaa
     2521 gcgctatggg ctgaatacaa accatcggat cgtggtgtgc agcgagaata gcttgcagtt
     2581 cttcatgccc gtgttgggtg ccctgttcat cggtgtggct gtggccccag ctaacgacat
     2641 ctacaacgag cgcgagctgc tgaacagcat gggcatcagc cagcccaccg tcgtattcgt
     2701 gagcaagaaa gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa
     2761 gatcatcatc atggatagca agaccgacta ccagggcttc caaagcatgt acaccttcgt
     2821 gacttcccat ttgccacccg gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg
     2881 ggacaaaacc atcgccctga tcatgaacag tagtggcagt accggattgc ccaagggcgt
     2941 agccctaccg caccgcaccg cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg
     3001 caaccagatc atccccgaca ccgctatcct cagcgtggtg ccatttcacc acggcttcgg
     3061 catgttcacc acgctgggct acttgatctg cggctttcgg gtcgtgctca tgtaccgctt
     3121 cgaggaggag ctattcttgc gcagcttgca agactataag attcaatctg ccctgctggt
     3181 gcccacacta tttagcttct tcgctaagag cactctcatc gacaagtacg acctaagcaa
     3241 cttgcacgag atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc
     3301 caaacgcttc cacctaccag gcatccgcca gggctacggc ctgacagaaa caaccagcgc
     3361 cattctgatc acccccgaag gggacgacaa gcctggcgca gtaggcaagg tggtgccctt
     3421 cttcgaggct aaggtggtgg acttggacac cggtaagaca ctgggtgtga accagcgcgg
     3481 cgagctgtgc gtccgtggcc ccatgatcat gagcggctac gttaacaacc ccgaggctac
     3541 aaacgctctc atcgacaagg acggctggct gcacagcggc gacatcgcct actgggacga
     3601 ggacgagcac ttcttcatcg tggaccggct gaagagcctg atcaaataca agggctacca
     3661 ggtagcccca gccgaactgg agagcatcct gctgcaacac cccaacatct tcgacgccgg
     3721 ggtcgccggc ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga
     3781 acacggtaaa accatgaccg agaaggagat cgtggactat gtggccagcc aggttacaac
     3841 cgccaagaag ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg
     3901 caagttggac gcccgcaaga tccgcgagat tctcattaag gccaagaagg gcggcaagat
     3961 cgccgtgaat tctgcttgca agaactggtt cagtagctta agccactttg tgatccacct
     4021 taacagccac ggcttccctc ccgaggtgga ggagcaggcc gccggcaccc tgcccatgag
     4081 ctgcgcccag gagagcggca tggatagaca ccctgctgct tgcgccagcg ccaggatcaa
     4141 cgtctaactg cagtctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     4201 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4261 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4321 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     4381 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc
     4441 ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     4501 ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4561 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4621 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4681 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4741 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4801 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4861 gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta
     4921 ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg atgcagttgt
     4981 cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc tatgtactgc
     5041 gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca
     5101 gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag agcctgacat
     5161 actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa acagagctgc
     5221 ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc gacctcagtc
     5281 aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca acctggcggg
     5341 atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg atggacgaca
     5401 ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc gaagactgtc
     5461 ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg accgacaacg
     5521 gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt cagtacgagg
     5581 tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag actcgctact
     5641 tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac atgctgcgca
     5701 tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat gctgcctggg
     5761 ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt cgcacacaaa
     5821 tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg gccgacagcg
     5881 gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag
     5941 aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     6001 gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     6061 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     6121 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     6181 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6241 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6961 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7021 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7081 ggtctgacag cggccgctaa ccctttaccg gcgcgaagcg ctacgcgctt cgcgagaggt
     7141 atctccgtgt gcttctccat cagaagacgc gtgcgccgct tcgcaggcga cccgcatact
     7201 ttccgccccg agtgcgttac aaaaatgact gccttctggc gacaatacac ggtggacgtc
     7261 cagtaccacc cgcatatcag cttatccggt ggcaatctgg cactggacag ggaattctcg
     7321 caacaatccg aggccatgat ggtggcagga ccgctggccg cacatagctc aatcacggcc
     7381 acccagaaga gcagccccaa atgtgcgcgc aacacccagc acatgctcca catacagttc
     7441 tggcgccaca acgatgatgc gcaaaggggt gcattaccct aaatcccagc ctagttataa
     7501 attattgaag cccaggcgac caggggtcgc cgcgcttttc ctccccaaac gcgacgataa
     7561 agaccagcgt tgccaaatgt aacttatgta taacccaaaa tattgcgcat cgataaggtt
     7621 tgccaaaaca cccgaaagta cacacacaaa aaaacagcaa caagacgctc actagacatt
     7681 caccccttcc cccacccccg aaaacaaaac aacttgacac aggggaaaca ccaggggcgg
     7741 cggaggttgt caatagtgtc cagtatttcg ttagacgcgg gttcttggac ccgatgtccc
     7801 aggtcattaa agtctcaaat gggattaaag gatcatagtt cccaggttta atactccaag
     7861 ctatcccaga acaggacccc ggcagaaccc cgctaacagc accaaatcca cttgcggtcc
     7921 cagaaaaggt cgccgaggtg gcaaggtgac tgaaaaggtc atagagagga caccggtccc
     7981 atttcccacg gtccaaaaat ccagcgcgcc ccaccggctt tccgagaact tcggcaaagc
     8041 taatttgcat gcgctaatcc ttttatgtgc ataaattatg tagatgagga gtcgcgcatg
     8101 cgcagaaaaa ttcagagcgc ccgggtgcac ggggtcacct ccaggtcacg ccgctaggtg
     8161 ggaccgtgag cgactcgaaa aattataatt tttggccatt ttatgggcgc cgccatcttg
     8221 aatttgctaa tcccccgtaa tcctctgccc ctctcccatt ggtccgccgg cccgtcaatc
     8281 aaagttttcc gagccgccat tggcccattc ggccgaccaa tcccgttcga gctaggcgac
     8341 cgcgccattc cattggacgc cccagccgtc aatcaaattc agaggcctcc cattggcccc
     8401 tatccctaga actcccaagc tgattggccc acagcgggaa ccaatcagcg attagagttt
     8461 tgttttgatt tttcctatat atatatataa tcctttaatc ctagcgcagc tgagtcatcg
     8521 cagcccctat tccagtaggt atacccagct gggtaatcca gtaggtatac ccaggtgggt
     8581 gaacccagct gggtataccc agctgcaatt ctataattaa acaaggtaga aaccaacggg
     8641 gtcctcaggt ggtatttccg gaagcattac caaataaggc aacctcagct gggaatacca
     8701 gcggactacc cccaactgta ttcaaccctc ctttgttttc cggaagtata tccatttatg
     8761 gaaatcagct gggtcactct actgggttat tctttataat agggcccgat gagtcatggg
     8821 gttgggattt ttctactagg tcgtttcggt ggatgggtgc caggattata ggggccctgt
     8881 ccacggggtt gttcggtggc g
//
