LOCUS       sequence-351441-        6945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KSHV ORF55 tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6945)
  AUTHORS   Alzhanova D, Corcoran K, Bailey AG, Long K, Taft-Benz S, Graham RL, 
            Broussard GS, Heise M, Neumann G, Halfmann P, Kawaoka Y, Baric RS, 
            Damania B, Dittmer DP
  TITLE     Novel modulators of p53-signaling encoded by unknown genes of 
            emerging viruses.
  JOURNAL   PLoS Pathog. 2021 Jan 7;17(1):e1009033. doi: 
            10.1371/journal.ppat.1009033. eCollection 2021 Jan.
  PUBMED    33411764
REFERENCE   2  (bases 1 to 6945)
  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: "PLoS 
            Pathog."; date: "2021-01-7"; volume: "17(1):e1009033. doi"; pages: "
            10.1371/journal.ppat.1009033. eCollection 2021 Jan"
COMMENT     Sequence Label: pOME0192
FEATURES             Location/Qualifiers
     source          1..6945
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    914..938
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1657..1722
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(1758..1782)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1793..1795,1796..1798,1799..2512)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1826..1854)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2449..2469
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2534..2551)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2540..2764
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2810..3238
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2897..2916)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3107..3128
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3247..3267)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3252..3581
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3432..3567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3494..3513
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3648..4442
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3702..3721)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4312..4331
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4618..4739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4655..4674)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4709..4728
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4788..4804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4788..4804)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4801..4823)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4812..4828
                     /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(4836..4842,4843..4860,4861..4866))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4881..4902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5019..5036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5190..5778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5270..5289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5949..6740,6741..6809))
                     /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     6572..6591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6810..6914)
                     /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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttgat caaacaagtt tgtacaaaaa agcaggctct ttaaaggaac caattcagtc
      961 gacggtaccg ccaccatgtc gtctccatgg tacacctgga cgtgttgcgg tataaacctt
     1021 tttggacgcg gcaaccacgc ttacaagcgc ctcggtgacc ctctggaagg ctgtccagag
     1081 cgctggcggc aagagataga tttgggcctc ccccccggag tgtgtctggg tgacgttgtc
     1141 cagtcgaacc ttggcacgac ggcgctgcac cagacctacc ttctggctgt acagtcaaac
     1201 aagatcacag actacctgaa gcgttttgac gtggctaaaa ttccggcggg ctgccaggag
     1261 accgtgaaaa cccaggtaaa aaaactgcag tctattcaga acgtcgtgtg gaacacaatg
     1321 ctggccttgg cggtcggcga gattaccgtg gacgacagcg cgctccagtc cctcttaaac
     1381 aaaagggccg gggaatgcgt ttcgctcatg gagatggaaa aactggcgac tgctatggcg
     1441 tcagacgact cggtaatatg ggcctctgag atcagccact ccttgtcgga acccacttct
     1501 gtattgcctc tcacgcctgc cgttacgcgc cagccggagg caactttacc caagccaccc
     1561 accgaagacc cgtccgtttc cgcgatgcat tcgtcaatcc ctccccggcc ttcgtcaacc
     1621 ctggaagaga cgacagaaag cgcgataggt tcgacagact acaaagacca tgacggtgat
     1681 tataaagatc atgatatcga ttacaaggat gacgatgaca agtagtaact cgaggcggcc
     1741 gcactcgaga tatctagacc cagctttctt gtacaaagtg gttcgatcta gaatggctag
     1801 caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
     1861 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgctacat acggaaagct
     1921 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac
     1981 tactttctct tatggtgttc aatgcttttc ccgttatccg gatcatatga aacggcatga
     2041 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga
     2101 tgacgggaac tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg
     2161 tatcgagtta aaaggtattg attttaaaga agatggaaac attctcggac acaaactcga
     2221 gtacaactat aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa
     2281 agctaacttc aaaattcgcc acaacattga agatggatcc gttcaactag cagaccatta
     2341 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
     2401 gacacaatct gccctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga
     2461 gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat aatgaattaa
     2521 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2581 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2641 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2701 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2761 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     2821 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2881 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2941 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3001 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3061 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3121 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3181 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3241 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3301 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3361 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3421 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3481 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3541 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3601 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3661 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3721 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3781 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3841 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3901 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3961 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4021 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4081 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4141 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4201 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4261 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4321 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4381 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4441 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4501 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4561 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4621 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4681 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4741 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4801 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4861 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4921 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4981 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5041 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5101 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5161 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5221 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5281 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5341 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5401 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5461 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5521 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5581 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5641 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5701 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5761 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5821 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5881 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5941 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6001 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6061 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6121 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6181 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6241 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6301 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6361 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6421 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6481 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6541 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6601 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6661 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6721 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6781 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6841 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6901 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
