LOCUS       sequence-256936-        7391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of KSHV ORF30 with a C-terminal 
            2xStrep tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7391)
  AUTHORS   Didychuk AL, Castaneda AF, Kushnir LO, Huang CJ, Glaunsinger BA
  TITLE     Conserved CxnC motifs in Kaposi's sarcoma-associated herpesvirus 
            ORF66 are required for viral late gene expression and are essential 
            for its interaction with ORF34.
  JOURNAL   J Virol. 2019 Oct 2. pii: JVI.01299-19. doi: 10.1128/JVI.01299-19.
  PUBMED    31578296
REFERENCE   2  (bases 1 to 7391)
  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. 
            2019 Oct 2. pii: JVI.01299-19. doi: 10.1128/JVI.01299-19."
COMMENT     Sequence Label: pLJM1-zeo-ORF30-2xStrep
FEATURES             Location/Qualifiers
     source          1..7391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     285..305
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             632..655
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             692..715
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    768..885
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        934..1444
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1363..1381
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1453..1827
                     /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"
     LTR             1899..2132
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2204..2325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2241..2260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2295..2314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2365..2500
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2520..2539)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2521..2539)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2549..2566)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2549..2565)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2558..2580)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2707..3162
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2794..2813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3004..3025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3188..3292
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3293..3361,3362..4153)
                     /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     complement(3511..3530)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4324..4912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4813..4832
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5066..5083
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5200..5221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5236..5241,5242..5259,5260..5266)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5274..5290
                     /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)."
     primer_bind     5279..5301
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5298..5314
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5298..5314
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5333..5353
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5335..5353
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5381..5607
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5608..5788
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5835..5960
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6453..6686
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6871..6915
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     7213..7233
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgctaccgg atgggtgagc cagtggatcc tggacatgtg gtgaatgaga aagattttga
      421 ggagtgtgaa caatttttca gtcaacccct tagggagcaa gtggtcgcgg gggtcagggc
      481 actcgacggc ctcggtctcg ctgactctct atgtcacaaa acagaaagac tctgcctgct
      541 gatggacctg gtgggcacgg agtgctttgc gagggtgtgc cgcctagaca ccggtgcgaa
      601 aggagggcgg ccgctcgagg gaggcggtgg atggagccac ccccagttcg agaagggcgg
      661 cggcagcggc ggcggcagcg gcggcggcag ctggagccac ccccagttcg agaaggggcc
      721 cgtttaaaat tctcgacctc gagacaaatg gcagtattca tccacaattt taaaagaaaa
      781 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
      841 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
      901 gacagcagag atccactttg gccgcggctc gagggggttg gggttgcgcc ttttccaagg
      961 cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg
     1021 cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc
     1081 cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc
     1141 cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct
     1201 cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa
     1261 tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc
     1321 ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa
     1381 gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc
     1441 ccagggggat ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     1501 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     1561 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     1621 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     1681 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     1741 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     1801 cacttcgtgg ccgaggagca ggactgaggt acctttaaga ccaatgactt acaaggcagc
     1861 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     1921 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     1981 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     2041 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     2101 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     2161 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     2221 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2281 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     2341 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2401 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2461 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     2521 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     2581 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2641 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2701 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2881 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3001 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3061 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3121 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     3181 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3241 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3301 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3361 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3421 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3481 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3541 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3601 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3661 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3721 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3781 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3841 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3901 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3961 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4021 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4081 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4141 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4201 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4261 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4321 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4381 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4441 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4501 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4561 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4621 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4681 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4741 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4801 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4861 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4921 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4981 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5041 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5101 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5161 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5221 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5281 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     5341 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     5401 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     5461 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     5521 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     5581 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     5641 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5701 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5761 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     5821 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     5881 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     5941 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     6001 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     6061 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     6121 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     6181 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     6241 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     6301 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     6361 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     6421 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     6481 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     6541 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     6601 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     6661 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     6721 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     6781 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     6841 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     6901 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     6961 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     7021 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     7081 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     7141 tccattcgat tagtgaacgg atctcgacgg tatcgatcac gagactagcc tcgagcggcc
     7201 gcccccttca ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     7261 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
     7321 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     7381 atggactatc a
//
