LOCUS       sequence-257145-        5998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase reporter with KSHV K8.1 late gene promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5998)
  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 5998)
  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: K8.1 Pr pGL4.16
FEATURES             Location/Qualifiers
     source          1..5998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      72..81
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             78..1727
                     /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             1734..1781
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             1785..1904
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    1953..2074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1964..1983)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2018..2037
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2268..2625
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2268..2459
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2291..2311)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2476..2611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2538..2557
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2656..3693
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3672..3680
                     /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    3717..3765
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3921..3938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4092..4680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4172..4191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4880..5671,5672..5740))
                     /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    5845..5893
                     /note="synthetic polyadenylation signal"
     misc_feature    5907..5998
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5947..5966
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtaccc cggcagcaat attaaaggga ccaagcttgg caatccggta
       61 ctgttggtaa agccaccatg gaagatgcca aaaacattaa gaagggccca gcgccattct
      121 acccactcga agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc
      181 tggtgcccgg caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg
      241 agtacttcga gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa
      301 accatcggat cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg
      361 ccctgttcat cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc
      421 tgaacagcat gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa
      481 agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca
      541 agaccgacta ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg
      601 gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga
      661 tcatgaacag tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg
      721 cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca
      781 ccgctatcct cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct
      841 acttgatctg cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc
      901 gcagcttgca agactataag attcaatctg ccctgctggt gcccacacta tttagcttct
      961 tcgctaagag cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg
     1021 gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag
     1081 gcatccgcca gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag
     1141 gggacgacaa gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg
     1201 acttggacac cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc
     1261 ccatgatcat gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg
     1321 acggctggct gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg
     1381 tggaccggct gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg
     1441 agagcatcct gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg
     1501 acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accatgaccg
     1561 agaaggagat cgtggactat gtggccagcc aggttacaac cgccaagaag ctgcgcggtg
     1621 gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga
     1681 tccgcgagat tctcattaag gccaagaagg gcggcaagat cgccgtgaat tctgcttgca
     1741 agaactggtt cagtagctta agccactttg tgatccacct taacagccac ggcttccctc
     1801 ccgaggtgga ggagcaggcc gccggcaccc tgcccatgag ctgcgcccag gagagcggca
     1861 tggatagaca ccctgctgct tgcgccagcg ccaggatcaa cgtctaactg cagtctagag
     1921 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     1981 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2041 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2101 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2161 tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc ttccgctgat ctgcgcagca
     2221 ccatggcctg aaataacctc tgaaagagga acttggttag ctaccttctg aggcggaaag
     2281 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2341 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2401 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2461 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2521 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     2581 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cgattcttct
     2641 gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga aaaatttctc
     2701 atcgagaagt tcgacagtgt gagcgacctg atgcagttgt cggagggcga agagagccga
     2761 gccttcagct tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag ctgcgctgat
     2821 ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact acccatcccc
     2881 gaagtgttgg acatcggcga gttcagcgag agcctgacat actgcatcag tagacgcgcc
     2941 caaggcgtta ctctccaaga cctccccgaa acagagctgc ctgctgtgtt acagcctgtc
     3001 gccgaagcta tggatgctat tgccgccgcc gacctcagtc aaaccagcgg cttcggccca
     3061 ttcgggcccc aaggcatcgg ccagtacaca acctggcggg atttcatttg cgccattgct
     3121 gatccccatg tctaccactg gcagaccgtg atggacgaca ccgtgtccgc cagcgtagct
     3181 caagccctgg acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg ccacctcgtc
     3241 catgccgact tcggcagcaa caacgtcctg accgacaacg gccgcatcac cgccgtaatc
     3301 gactggtccg aagctatgtt cggggacagt cagtacgagg tggccaacat cttcttctgg
     3361 cggccctggc tggcttgcat ggagcagcag actcgctact tcgagcgccg gcatcccgag
     3421 ctggccggca gccctcgtct gcgagcctac atgctgcgca tcggcctgga tcagctctac
     3481 cagagcctcg tggacggcaa cttcgacgat gctgcctggg ctcaaggccg ctgcgatgcc
     3541 atcgtccgca gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg gagcgcagcc
     3601 gtatggaccg acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg gcccagtaca
     3661 cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag aaccggtcat ggccgcaata
     3721 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgttcga actagatgct
     3781 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     3841 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     3901 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3961 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4021 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4081 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4141 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4201 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4261 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4321 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4381 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4441 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4501 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4561 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4621 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4801 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag cggccgcaaa
     4861 tgctaaacca ctgcagtggt taccagtgct tgatcagtga ggcaccgatc tcagcgatct
     4921 gcctatttcg ttcgtccata gtggcctgac tccccgtcgt gtagatcact acgattcgtg
     4981 agggcttacc atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt tcaccggccc
     5041 ccgatttgtc agcaatgaac cagccagcag ggagggccga gcgaagaagt ggtcctgcta
     5101 ctttgtccgc ctccatccag tctatgagct gctgtcgtga tgctagagta agaagttcgc
     5161 cagtgagtag tttccgaaga gttgtggcca ttgctactgg catcgtggta tcacgctcgt
     5221 cgttcggtat ggcttcgttc aactctggtt cccagcggtc aagccgggtc acatgatcac
     5281 ccatattatg aagaaatgca gtcagctcct tagggcctcc gatcgttgtc agaagtaagt
     5341 tggccgcggt gttgtcgctc atggtaatgg cagcactaca caattctctt accgtcatgc
     5401 catccgtaag atgcttttcc gtgaccggcg agtactcaac caagtcgttt tgtgagtagt
     5461 gtatacggcg accaagctgc tcttgcccgg cgtctatacg ggacaacacc gcgccacata
     5521 gcagtacttt gaaagtgctc atcatcggga atcgttcttc ggggcggaaa gactcaagga
     5581 tcttgccgct attgagatcc agttcgatat agcccactct tgcacccagt tgatcttcag
     5641 catcttttac tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa aatgccgcaa
     5701 agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt tttcaatatt
     5761 attgaagcat ttatcagggt tactagtacg tctctcaagg ataagtaagt aatattaagg
     5821 tacgggaggt attggacagg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     5881 ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa
     5941 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctct
//
