LOCUS       sequence-257262-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase reporter with KSHV ORF57 early gene promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  AUTHORS   Castaneda AF, Glaunsinger BA
  TITLE     The interaction between ORF18 and ORF30 is required for late gene 
            expression in Kaposi's sarcoma-associated herpesvirus.
  JOURNAL   J Virol. 2018 Oct 10. pii: JVI.01488-18. doi: 10.1128/JVI.01488-18.
  PUBMED    30305361
REFERENCE   2  (bases 1 to 6011)
  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. 
            2018 Oct 10. pii: JVI.01488-18. doi: 10.1128/JVI.01488-18."
COMMENT     Sequence Label: ORF57 Pr pGL4.16
FEATURES             Location/Qualifiers
     source          1..6011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      85..94
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             91..1740
                     /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             1747..1794
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             1798..1917
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    1966..2087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1977..1996)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2031..2050
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2281..2638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2281..2472
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2304..2324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2489..2624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2551..2570
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2669..3706
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3685..3693
                     /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    3730..3778
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3934..3951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4105..4693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4185..4204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4893..5684,5685..5753))
                     /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    5858..5906
                     /note="synthetic polyadenylation signal"
     misc_feature    5920..6011
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5960..5979
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtaccg gccagttaat cccactatat aacctggctg ccaggaagct
       61 tggcaatccg gtactgttgg taaagccacc atggaagatg ccaaaaacat taagaagggc
      121 ccagcgccat tctacccact cgaagacggg accgccggcg agcagctgca caaagccatg
      181 aagcgctacg ccctggtgcc cggcaccatc gcctttaccg acgcacatat cgaggtggac
      241 attacctacg ccgagtactt cgagatgagc gttcggctgg cagaagctat gaagcgctat
      301 gggctgaata caaaccatcg gatcgtggtg tgcagcgaga atagcttgca gttcttcatg
      361 cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc cagctaacga catctacaac
      421 gagcgcgagc tgctgaacag catgggcatc agccagccca ccgtcgtatt cgtgagcaag
      481 aaagggctgc aaaagatcct caacgtgcaa aagaagctac cgatcataca aaagatcatc
      541 atcatggata gcaagaccga ctaccagggc ttccaaagca tgtacacctt cgtgacttcc
      601 catttgccac ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa
      661 accatcgccc tgatcatgaa cagtagtggc agtaccggat tgcccaaggg cgtagcccta
      721 ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg accccatctt cggcaaccag
      781 atcatccccg acaccgctat cctcagcgtg gtgccatttc accacggctt cggcatgttc
      841 accacgctgg gctacttgat ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag
      901 gagctattct tgcgcagctt gcaagactat aagattcaat ctgccctgct ggtgcccaca
      961 ctatttagct tcttcgctaa gagcactctc atcgacaagt acgacctaag caacttgcac
     1021 gagatcgcca gcggcggggc gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc
     1081 ttccacctac caggcatccg ccagggctac ggcctgacag aaacaaccag cgccattctg
     1141 atcacccccg aaggggacga caagcctggc gcagtaggca aggtggtgcc cttcttcgag
     1201 gctaaggtgg tggacttgga caccggtaag acactgggtg tgaaccagcg cggcgagctg
     1261 tgcgtccgtg gccccatgat catgagcggc tacgttaaca accccgaggc tacaaacgct
     1321 ctcatcgaca aggacggctg gctgcacagc ggcgacatcg cctactggga cgaggacgag
     1381 cacttcttca tcgtggaccg gctgaagagc ctgatcaaat acaagggcta ccaggtagcc
     1441 ccagccgaac tggagagcat cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc
     1501 ggcctgcccg acgacgatgc cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt
     1561 aaaaccatga ccgagaagga gatcgtggac tatgtggcca gccaggttac aaccgccaag
     1621 aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta aaggactgac cggcaagttg
     1681 gacgcccgca agatccgcga gattctcatt aaggccaaga agggcggcaa gatcgccgtg
     1741 aattctgctt gcaagaactg gttcagtagc ttaagccact ttgtgatcca ccttaacagc
     1801 cacggcttcc ctcccgaggt ggaggagcag gccgccggca ccctgcccat gagctgcgcc
     1861 caggagagcg gcatggatag acaccctgct gcttgcgcca gcgccaggat caacgtctaa
     1921 ctgcagtcta gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg
     1981 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2041 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2101 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     2161 acctctacaa atgtggtaaa atcgataagg atccgtttgc gtattgggcg ctcttccgct
     2221 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt tagctacctt
     2281 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2341 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2401 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2461 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2521 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     2581 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2641 gctcgattct tctgacacta gcgccaccat gaagaagccc gaactcaccg ctaccagcgt
     2701 tgaaaaattt ctcatcgaga agttcgacag tgtgagcgac ctgatgcagt tgtcggaggg
     2761 cgaagagagc cgagccttca gcttcgatgt cggcggacgc ggctatgtac tgcgggtgaa
     2821 tagctgcgct gatggcttct acaaagaccg ctacgtgtac cgccacttcg ccagcgctgc
     2881 actacccatc cccgaagtgt tggacatcgg cgagttcagc gagagcctga catactgcat
     2941 cagtagacgc gcccaaggcg ttactctcca agacctcccc gaaacagagc tgcctgctgt
     3001 gttacagcct gtcgccgaag ctatggatgc tattgccgcc gccgacctca gtcaaaccag
     3061 cggcttcggc ccattcgggc cccaaggcat cggccagtac acaacctggc gggatttcat
     3121 ttgcgccatt gctgatcccc atgtctacca ctggcagacc gtgatggacg acaccgtgtc
     3181 cgccagcgta gctcaagccc tggacgaact gatgctgtgg gccgaagact gtcccgaggt
     3241 gcgccacctc gtccatgccg acttcggcag caacaacgtc ctgaccgaca acggccgcat
     3301 caccgccgta atcgactggt ccgaagctat gttcggggac agtcagtacg aggtggccaa
     3361 catcttcttc tggcggccct ggctggcttg catggagcag cagactcgct acttcgagcg
     3421 ccggcatccc gagctggccg gcagccctcg tctgcgagcc tacatgctgc gcatcggcct
     3481 ggatcagctc taccagagcc tcgtggacgg caacttcgac gatgctgcct gggctcaagg
     3541 ccgctgcgat gccatcgtcc gcagcggggc cggcaccgtc ggtcgcacac aaatcgctcg
     3601 ccggagcgca gccgtatgga ccgacggctg cgtcgaggtg ctggccgaca gcggcaaccg
     3661 ccggcccagt acacgaccgc gcgctaagga ggtaggtcga gtttaaactc tagaaccggt
     3721 catggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgtt
     3781 cgaactagat gctgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     3841 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     3901 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3961 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4021 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4081 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4141 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4201 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4261 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4321 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4381 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4441 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4501 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4561 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4621 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4681 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4741 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4801 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4861 cagcggccgc aaatgctaaa ccactgcagt ggttaccagt gcttgatcag tgaggcaccg
     4921 atctcagcga tctgcctatt tcgttcgtcc atagtggcct gactccccgt cgtgtagatc
     4981 actacgattc gtgagggctt accatcaggc cccagcgcag caatgatgcc gcgagagccg
     5041 cgttcaccgg cccccgattt gtcagcaatg aaccagccag cagggagggc cgagcgaaga
     5101 agtggtcctg ctactttgtc cgcctccatc cagtctatga gctgctgtcg tgatgctaga
     5161 gtaagaagtt cgccagtgag tagtttccga agagttgtgg ccattgctac tggcatcgtg
     5221 gtatcacgct cgtcgttcgg tatggcttcg ttcaactctg gttcccagcg gtcaagccgg
     5281 gtcacatgat cacccatatt atgaagaaat gcagtcagct ccttagggcc tccgatcgtt
     5341 gtcagaagta agttggccgc ggtgttgtcg ctcatggtaa tggcagcact acacaattct
     5401 cttaccgtca tgccatccgt aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg
     5461 ttttgtgagt agtgtatacg gcgaccaagc tgctcttgcc cggcgtctat acgggacaac
     5521 accgcgccac atagcagtac tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg
     5581 aaagactcaa ggatcttgcc gctattgaga tccagttcga tatagcccac tcttgcaccc
     5641 agttgatctt cagcatcttt tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag
     5701 caaaatgccg caaagaaggg aatgagtgcg acacgaaaat gttggatgct catactcgtc
     5761 ctttttcaat attattgaag catttatcag ggttactagt acgtctctca aggataagta
     5821 agtaatatta aggtacggga ggtattggac aggccgcaat aaaatatctt tattttcatt
     5881 acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa
     5941 caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag
     6001 aacatttctc t
//
