LOCUS       sequence-257147-        7389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KSHV ORF24 with Cterm 2xStrep tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7389)
  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 7389)
  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: ORF24-2xStrep pCDNA4
FEATURES             Location/Qualifiers
     source          1..7389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3288..3311
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3348..3371
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3397..3414)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3403..3627
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3673..4101
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3760..3779)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3970..3991
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4110..4130)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4115..4444
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4295..4430
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4357..4376
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4492..4539
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4558..4932
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5062..5183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5099..5118)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5153..5172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5232..5248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5232..5248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5245..5267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5256..5272
                     /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(5280..5286,5287..5304,5305..5310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5325..5346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5463..5480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5634..6222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5714..5733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6393..7184,7185..7253))
                     /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     7016..7035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7254..7358)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc catggcagcg ctcgagggcc
     1021 ccctactact gccaccgagc gcctccctga cgacgagtcc gcagaccacg tgttatcaag
     1081 cgacttggga atcacagctg gaaatattct gctgtctggc caccaactcg cacctgcagg
     1141 cagagctgac cttagaaggt cttgataaga tgatgcagcc cgagcccacc tttttcgcct
     1201 gcagagcgat acgcagacta ctcctggggg aacgcctcca cccttttata catcaagaag
     1261 ggactctttt gggaaaagtg ggtcgacggt acagcggcga aggtttaata attgacggtg
     1321 gtggagtgtt tacgcgcgga cagatagaca ccgacaacta cctacctgcg gtgggatcat
     1381 gggaacttac cgatgattgt gataaaccct gcgaattcag ggagctacgc tcgctgtatc
     1441 ttcccgcgct actaacgtgc accatatgtt acaaagccat gttcaggata gtgtgcaggt
     1501 acctggagtt ctgggagttc gaacagtgtt ttcatgcgtt tctggcggtg ttgccccata
     1561 gtctacaacc cacaatctat caaaattatt ttgcactcct ggagagcctg aagcatctct
     1621 cgttttcaat aatgccaccc gcatccccag acgcacagct acatttttta aagtttaaca
     1681 tcggcagctt catggccacg tgggggtggc acggagagct ggtctcgctg cgccgtgcca
     1741 tcgctcacaa cgtagagcga ctgcccaccg tgctgaagaa cctgtcgaaa cagagtaagc
     1801 accaggacgt caaggttaac ggacgggatc tggtgggctt tcagctggct ctaaaccagc
     1861 tcgtgtcccg tctgcacgta aaaatccaac gcaaggaccc cggaccaaag ccatacaggg
     1921 tggtcgtcag taccccagat tgtacctact atctagtgta tccgggcaca ccggccatct
     1981 acagactcgt catgtgtatg gcagtggcag actgcatcgg ccactcgtgc agcggactgc
     2041 acccctgcgc aaacttttta ggcacccacg agacaccgcg tctcctggcg gcgacgcttt
     2101 caagaatccg gtacgcgccg aaagaccggc gagcagccat gaagggaaat ttgcaggcgt
     2161 gcttccaacg atacgcggcc acggacgcgc ggactctggg cagctccaca gtgtcagaca
     2221 tgctggaacc cacaaaacac gtcagtttgg aaaacttcaa gatcaccata ttcaacacca
     2281 acatggtgat taacactaag ataagctgcc acgttcctaa caccctgcaa aagactattt
     2341 taaacatccc cagattgacc aacaattttg ttatacgaaa gtactccgta aaggaacctt
     2401 cttttaccat aagcgtgttt ttttccgaca acatgtgtca aggcaccgca ataaacatca
     2461 acatcagtgg ggacatgctg cactttctct tcgcaatggg tacgctgaaa tgctttctgc
     2521 caatcaggca catatttcct gtatcgatag caaattggaa ctccacgttg gacctgcacg
     2581 gactggaaaa ccagtacatg gtgagaatgg ggcgaaaaaa cgtattttgg accacaaact
     2641 ttccatctgt ggtctccagc aaggatgggc taaacgtgtc ctggtttaag gccgcgacag
     2701 ccacgatttc taaagtgtac gggcagcctc ttgtggaaca gattcgccac gagctggcgc
     2761 ccattctcac ggaccagcac gcgcgcatcg acggaaacaa aaatagaata ttctccctac
     2821 ttgagcacag aaaccgttcc caaatacaga cgctacacaa aaggttcctg gagtgtctgg
     2881 tggaatgctg ttcgtttctc aggcttgacg tggcttgcat taggcgagcc gccgcccggg
     2941 gcctgtttga cttctcaaag aagataatca gtcacactaa aagcaaacac gagtgcgcag
     3001 tactgggata taaaaagtgt aacctaatcc cgaaaatcta tgcccgaaac aagaagacca
     3061 ggctagacga gttgggccgc aatgcaaact tcatttcgtt cgtcgccacc acgggtcatc
     3121 ggttcgccgc tctaaagcca caaattgtcc gtcacgccat tcgcaaacta ggcctgcact
     3181 ggcgccaccg aacggccgcg tccaacgagc agacaccgcc agccgatccc cgcgtacgtt
     3241 gcgtccgtcc gctggtctca gggcggccgc tcgagggagg cggtggatgg agccaccccc
     3301 agttcgagaa gggcggcggc agcggcggcg gcagcggcgg cggcagctgg agccaccccc
     3361 agttcgagaa ggggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     3421 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3481 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3541 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3601 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3661 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3721 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3781 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3841 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3901 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3961 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4021 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4081 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4201 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4261 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4321 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4381 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4441 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     4501 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     4561 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     4621 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     4681 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     4741 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     4801 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     4861 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     4921 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4981 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5041 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5101 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5161 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5221 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5281 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5341 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5401 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5461 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5821 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6061 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7381 cctgacgtc
//
