LOCUS       sequence-257146-        6087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KSHV ORF34 with Nterm 2xStrep tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6087)
  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 6087)
  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: 2xStrep-ORF34 pCDNA4
FEATURES             Location/Qualifiers
     source          1..6087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    819..837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    840..858
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     868..892
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             990..1013
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1050..1073
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2094..2111)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2100..2324
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2370..2798
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2457..2476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2667..2688
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2807..2827)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2812..3141
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2992..3127
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3054..3073
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3189..3236
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3255..3629
                     /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    3759..3880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3796..3815)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3850..3869
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3929..3945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3929..3945)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3942..3964)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3953..3969
                     /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(3977..3983,3984..4001,4002..4007))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4022..4043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4160..4177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4331..4919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4411..4430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5090..5881,5882..5950))
                     /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     5713..5732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5951..6055)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct
      841 ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag
      901 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac
      961 tctagcgttt aaacttaagc ttaccaatgt ggagccaccc ccagttcgag aagggcggcg
     1021 gcagcggcgg cggcagcggc ggcggcagct ggagccaccc ccagttcgag aagggggcgg
     1081 cctttgcttt gagctcgctc gtgtccgagg gtgacccgga ggtgaccagt aggtacgtca
     1141 agggcgtaca acttgccctg gaccttagcg agaacacacc tggacaattt aagttgatag
     1201 aaactcccct gaacagcttc ctcttggttt ccaacgtgat gcccgaggtc cagccaatct
     1261 gcagtggccg gccggccttg cggccagact ttagtaatct ccacttgcct agactggaga
     1321 agctccagag agtcctcggg cagggtttcg gggcggcggg tgaggaaatc gcactggacc
     1381 cgtctcacgt agaaacacac gaaaagggcc aggtgttcta caaccactat gctaccgagg
     1441 agtggacgtg ggctttgact ctgaataagg atgcgctcct tcgggaggct gtagatggcc
     1501 tgtgtgaccc cggaacttgg aagggtcttc ttcctgacga cccccttccg ttgctatggc
     1561 tgctgttcaa cggacccgcc tctttttgtc gggccgactg ttgcctgtac aagcagcact
     1621 gcggttaccc gggcccggtg ctacttccag gtcacatgta cgctcccaaa cgggatcttt
     1681 tgtcgttcgt taatcatgcc ctgaagtaca ccaagtttct atacggagat ttttccggga
     1741 catgggcggc ggcttgccgc ccgccattcg ctacttctcg gatacaaagg gtagtgagtc
     1801 agatgaaaat catagatgct tccgacactt acatttccca cacctgcctc ttgtgtcaca
     1861 tatatcagca aaatagcata attgcgggtc aggggaccca cgtgggtgga atcctactgt
     1921 tgagtggaaa agggacccag tatataacag gcaatgttca gacccaaagg tgtccaacta
     1981 cgggcgacta tctaatcatc ccatcgtatg acataccggc gatcatcacc atgatcaagg
     2041 agaatggact caaccaactc taactagagg gcccgtttaa acccgctgat cagcctcgac
     2101 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2161 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2221 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2281 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2341 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2401 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2461 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2521 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2581 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2641 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2701 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2761 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2821 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2881 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2941 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3001 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3061 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3121 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3181 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3241 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     3301 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     3361 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     3421 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     3481 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     3541 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     3601 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     3661 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3721 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3781 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3841 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3901 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3961 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4021 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4081 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4141 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4201 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4261 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4321 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4381 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4441 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4501 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4561 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4621 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4681 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4741 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4801 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4861 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4921 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4981 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5041 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5101 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5161 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5221 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5281 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5341 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5401 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5461 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5521 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5581 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5641 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5701 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5761 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5821 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5881 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5941 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6001 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6061 ttccccgaaa agtgccacct gacgtcg
//
