LOCUS       sequence-257152-        6075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly Luciferase reporter for K8.1 promoter from KSHV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6075)
  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 6075)
  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
FEATURES             Location/Qualifiers
     source          1..6075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      29..38
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             35..1684
                     /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             1691..1738
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             1742..1861
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    1910..2031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1921..1940)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1975..1994
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2225..2582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2225..2416
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2248..2268)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2433..2568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2495..2514
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2613..3650
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3629..3637
                     /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    3674..3722
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3878..3895)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4049..4637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4129..4148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4837..5628,5629..5697))
                     /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    5802..5850
                     /note="synthetic polyadenylation signal"
     misc_feature    5864..5955
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5904..5923
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 agcttggcaa tccggtactg ttggtaaagc caccatggaa gatgccaaaa acattaagaa
       61 gggcccagcg ccattctacc cactcgaaga cgggaccgcc ggcgagcagc tgcacaaagc
      121 catgaagcgc tacgccctgg tgcccggcac catcgccttt accgacgcac atatcgaggt
      181 ggacattacc tacgccgagt acttcgagat gagcgttcgg ctggcagaag ctatgaagcg
      241 ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc gagaatagct tgcagttctt
      301 catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta acgacatcta
      361 caacgagcgc gagctgctga acagcatggg catcagccag cccaccgtcg tattcgtgag
      421 caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca tacaaaagat
      481 catcatcatg gatagcaaga ccgactacca gggcttccaa agcatgtaca ccttcgtgac
      541 ttcccatttg ccacccggct tcaacgagta cgacttcgtg cccgagagct tcgaccggga
      601 caaaaccatc gccctgatca tgaacagtag tggcagtacc ggattgccca agggcgtagc
      661 cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca tcttcggcaa
      721 ccagatcatc cccgacaccg ctatcctcag cgtggtgcca tttcaccacg gcttcggcat
      781 gttcaccacg ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt accgcttcga
      841 ggaggagcta ttcttgcgca gcttgcaaga ctataagatt caatctgccc tgctggtgcc
      901 cacactattt agcttcttcg ctaagagcac tctcatcgac aagtacgacc taagcaactt
      961 gcacgagatc gccagcggcg gggcgccgct cagcaaggag gtaggtgagg ccgtggccaa
     1021 acgcttccac ctaccaggca tccgccaggg ctacggcctg acagaaacaa ccagcgccat
     1081 tctgatcacc cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg tgcccttctt
     1141 cgaggctaag gtggtggact tggacaccgg taagacactg ggtgtgaacc agcgcggcga
     1201 gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt aacaaccccg aggctacaaa
     1261 cgctctcatc gacaaggacg gctggctgca cagcggcgac atcgcctact gggacgagga
     1321 cgagcacttc ttcatcgtgg accggctgaa gagcctgatc aaatacaagg gctaccaggt
     1381 agccccagcc gaactggaga gcatcctgct gcaacacccc aacatcttcg acgccggggt
     1441 cgccggcctg cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg tgctggaaca
     1501 cggtaaaacc atgaccgaga aggagatcgt ggactatgtg gccagccagg ttacaaccgc
     1561 caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac tgaccggcaa
     1621 gttggacgcc cgcaagatcc gcgagattct cattaaggcc aagaagggcg gcaagatcgc
     1681 cgtgaattct gcttgcaaga actggttcag tagcttaagc cactttgtga tccaccttaa
     1741 cagccacggc ttccctcccg aggtggagga gcaggccgcc ggcaccctgc ccatgagctg
     1801 cgcccaggag agcggcatgg atagacaccc tgctgcttgc gccagcgcca ggatcaacgt
     1861 ctaactgcag tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
     1921 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1981 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2041 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2101 taaaacctct acaaatgtgg taaaatcgat aaggatccgt ttgcgtattg ggcgctcttc
     2161 cgctgatctg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttagcta
     2221 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2281 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2341 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2401 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2461 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2521 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2581 aaaagctcga ttcttctgac actagcgcca ccatgaagaa gcccgaactc accgctacca
     2641 gcgttgaaaa atttctcatc gagaagttcg acagtgtgag cgacctgatg cagttgtcgg
     2701 agggcgaaga gagccgagcc ttcagcttcg atgtcggcgg acgcggctat gtactgcggg
     2761 tgaatagctg cgctgatggc ttctacaaag accgctacgt gtaccgccac ttcgccagcg
     2821 ctgcactacc catccccgaa gtgttggaca tcggcgagtt cagcgagagc ctgacatact
     2881 gcatcagtag acgcgcccaa ggcgttactc tccaagacct ccccgaaaca gagctgcctg
     2941 ctgtgttaca gcctgtcgcc gaagctatgg atgctattgc cgccgccgac ctcagtcaaa
     3001 ccagcggctt cggcccattc gggccccaag gcatcggcca gtacacaacc tggcgggatt
     3061 tcatttgcgc cattgctgat ccccatgtct accactggca gaccgtgatg gacgacaccg
     3121 tgtccgccag cgtagctcaa gccctggacg aactgatgct gtgggccgaa gactgtcccg
     3181 aggtgcgcca cctcgtccat gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc
     3241 gcatcaccgc cgtaatcgac tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg
     3301 ccaacatctt cttctggcgg ccctggctgg cttgcatgga gcagcagact cgctacttcg
     3361 agcgccggca tcccgagctg gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg
     3421 gcctggatca gctctaccag agcctcgtgg acggcaactt cgacgatgct gcctgggctc
     3481 aaggccgctg cgatgccatc gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg
     3541 ctcgccggag cgcagccgta tggaccgacg gctgcgtcga ggtgctggcc gacagcggca
     3601 accgccggcc cagtacacga ccgcgcgcta aggaggtagg tcgagtttaa actctagaac
     3661 cggtcatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     3721 tgttcgaact agatgctgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     3781 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     3841 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3901 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3961 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4021 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4081 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4141 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4201 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4261 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4321 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4381 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4441 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4501 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4561 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4621 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4681 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4741 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4801 ctgacagcgg ccgcaaatgc taaaccactg cagtggttac cagtgcttga tcagtgaggc
     4861 accgatctca gcgatctgcc tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta
     4921 gatcactacg attcgtgagg gcttaccatc aggccccagc gcagcaatga tgccgcgaga
     4981 gccgcgttca ccggcccccg atttgtcagc aatgaaccag ccagcaggga gggccgagcg
     5041 aagaagtggt cctgctactt tgtccgcctc catccagtct atgagctgct gtcgtgatgc
     5101 tagagtaaga agttcgccag tgagtagttt ccgaagagtt gtggccattg ctactggcat
     5161 cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac tctggttccc agcggtcaag
     5221 ccgggtcaca tgatcaccca tattatgaag aaatgcagtc agctccttag ggcctccgat
     5281 cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa
     5341 ttctcttacc gtcatgccat ccgtaagatg cttttccgtg accggcgagt actcaaccaa
     5401 gtcgttttgt gagtagtgta tacggcgacc aagctgctct tgcccggcgt ctatacggga
     5461 caacaccgcg ccacatagca gtactttgaa agtgctcatc atcgggaatc gttcttcggg
     5521 gcggaaagac tcaaggatct tgccgctatt gagatccagt tcgatatagc ccactcttgc
     5581 acccagttga tcttcagcat cttttacttt caccagcgtt tcggggtgtg caaaaacagg
     5641 caagcaaaat gccgcaaaga agggaatgag tgcgacacga aaatgttgga tgctcatact
     5701 cgtccttttt caatattatt gaagcattta tcagggttac tagtacgtct ctcaaggata
     5761 agtaagtaat attaaggtac gggaggtatt ggacaggccg caataaaata tctttatttt
     5821 cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc
     5881 aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg
     5941 ccagaacatt tctctggcct aactggccgg tacctgtgcc gtcgtccggg agaaccatgc
     6001 cagactttgt gtggtaagaa ggaattgtta tccggcagca atattaaagg gaccgaagtt
     6061 aatcccttaa tccta
//
