LOCUS       sequence-351327-        6699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KSHV ORF45 tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6699)
  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 6699)
  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: pOME0016
FEATURES             Location/Qualifiers
     source          1..6699
                     /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"
     regulatory      913..922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2140..2205
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2252..2270)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2253..2270)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2290..2307)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2296..2520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2566..2994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2653..2672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2863..2884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3003..3023)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3008..3337
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3188..3323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3250..3269
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3404..4198
                     /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(3458..3477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4068..4087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4372..4493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4409..4428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4463..4482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4542..4558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4542..4558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4555..4577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4566..4582
                     /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(4590..4596,4597..4614,4615..4620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4635..4656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4773..4790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4944..5532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5024..5043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5703..6494,6495..6563))
                     /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     6326..6345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6564..6668)
                     /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 gcttggtacc
      901 gagctcggat ccgccaccat ggcgatgttt gtgaggacct cgtctagcac acacgatgaa
      961 gagagaatgc ttccaattga aggagcgcct cgcagacgac cccccgtgaa gttcatattc
     1021 ccacctccac ctctttcatc acttccagga tttggcaggc cgcgcggcta tgctggaccc
     1081 acggtgatag atatgtctgc cccagacgac gtcttcgccg aggacacgcc atcgccgcca
     1141 gcaacccctc tggatctaca gatatccccg gatcagtcga gcggcgaatc tgaatatgac
     1201 gaggatgagg aagatgaaga tgaagaagaa aatgacgatg ttcaggagga agacgagcca
     1261 gaggggtacc ctgcagactt ttttcaacct ttatctcact tgcgcccgag gcctctggcc
     1321 agacgggccc atacgcccaa accggtagca gtggtagcgg gccgcgtgcg cagttcaacg
     1381 gacacggcgg agtccgaggc gtccatggga tgggttagtc aggatgacgg attttcccct
     1441 gctgggctct caccttcaga cgacgagggg gttgctatcc tggaaccgat ggcggcatac
     1501 actgggaccg gggcatacgg actttcacct gcttccagaa atagtgtacc tggaacacaa
     1561 agttcaccat acagcgaccc tgatgaaggg ccctcgtggc gccccctgcg cgccgcaccc
     1621 accgcgatcg tcgacctgac atcggactct gatagcgatg acagttccaa ctctccggac
     1681 gtgaacaatg aggccgcgtt taccgacgcg cgccattttt cccaccagcc accctcgtcc
     1741 gaggaggacg gagaagacca aggggaagta ttgagtcaga gaatcgggct catggacgtg
     1801 ggccagaagc gcaaaaggca gtctaccgcc tcctctggta gcgaggatgt ggtgcgctgc
     1861 cagagacaac caaacttaag ccgcaaagca gtggcgtccg tgataattat atcctcgggg
     1921 agtgacacag acgaggagcc ctcgtccgcc gtgagcgtga tcgtgtctcc gtcgagcaca
     1981 aagggtcacc tcccaaccca atctcccagt acttccgccc actcgatttc atcaggaagc
     2041 acaactaccg cggggtccag gtgcagcgac ccaacccgca tcctggcctc cacgccaccc
     2101 ctgtgtggaa acggtgcata taactggccg tggctggacg actacaaaga ccatgacggt
     2161 gattataaag atcatgatat cgattacaag gatgacgatg acaagtagta actcgaggcg
     2221 gccgctcgag catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc
     2281 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2341 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2401 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2461 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2521 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     2581 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2641 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2701 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2761 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2821 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2881 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2941 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3001 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3061 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3181 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3241 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3301 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3361 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3421 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3481 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3541 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     3601 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3661 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3721 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3781 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3841 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3901 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     3961 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4021 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4081 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4141 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4201 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4261 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4321 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4381 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4441 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4501 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4561 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4621 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4681 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4741 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4801 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4861 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4921 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4981 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5041 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5101 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5161 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5221 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5281 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5341 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5401 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5461 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5521 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5581 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5641 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5701 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5761 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5821 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5881 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5941 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6001 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6061 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6121 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6181 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6241 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6301 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6361 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6421 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6481 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6541 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6601 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6661 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
