LOCUS       sequence-351322-        6712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KSHV pre-miRNA K12-2wt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6712)
  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 6712)
  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: pOME0207
FEATURES             Location/Qualifiers
     source          1..6712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        370..573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      612..621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             618..1295
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(748..766)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(798..818)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1016..1035
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1210..1233
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     LTR             1415..1840
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(1943..1965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2101..2121)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2106..2435
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2286..2421
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2457..2476
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2557..2574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2728..3316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2808..2827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3487..4278,4279..4347))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4110..4129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4348..4452)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4520..4538
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4638..5017
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5018..5221
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5222..5397
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5460..5659
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5860..6276
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6206..6228
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6244..6260
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        6300..6548
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6300..6320
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6471..6490
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aattctagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      601 cgctaccggt cgccaccatg gcctcctccg aggacgtcat caaggagttc atgcgcttca
      661 aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
      721 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
      781 ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg tacgtgaagc
      841 accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc aagtgggagc
      901 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
      961 acggctcctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc gacggccccg
     1021 taatgcagaa gaagactatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg
     1081 gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa ggacggcggc cactacctgg
     1141 tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc tactactacg
     1201 tggactccaa gctggacatc acctcccaca acgaggacta caccatcgtg gagcagtacg
     1261 agcgcgccga gggccgccac cacctgttcc tgtagctcga gatatcagtg gtccaggctc
     1321 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat
     1381 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     1441 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     1501 agaagttcag atcaaggtca ggaacagatg gaacagggtc gaccctagag aaccatcaga
     1561 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     1621 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     1681 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     1741 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     1801 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
     1861 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc tgcctcgcgc
     1921 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1981 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2041 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtag atccggctgt
     2101 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2161 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2221 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2281 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2341 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2401 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tactggctta actatgcggc
     2461 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     2521 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2581 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2641 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3781 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4081 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     4141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4381 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4441 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4501 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattagc
     4561 ttggccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
     4621 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     4681 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     4741 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     4801 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     4861 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     4921 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     4981 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     5041 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     5101 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     5161 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     5221 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cttccgatag actgcgtcgc
     5281 ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggtctcgct
     5341 gttccttggg agggtctcct ctgagtgatt gactacccac gacgggggtc tttcatttgg
     5401 gggctcgtcc gggatttgga gacccctgcc cagggaccac cgacccacca ccgggaggta
     5461 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgtt tgatgttatg
     5521 cgcctgcgtc tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
     5581 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt
     5641 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg
     5701 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg
     5761 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt
     5821 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc
     5881 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     5941 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     6001 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     6061 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc
     6121 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     6181 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     6241 tccctttatc cagccctcac tccttctcta ggcgccggaa ttgaagatct gggcaggaag
     6301 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     6361 taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     6421 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     6481 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     6541 gaggatccga aagtccgtca accagattta gattaagatt ataatgggtc tacttcgcta
     6601 actgtagtcc gggtcgatct gagccattga agcaagcttc cagatcttcc agggctagag
     6661 ctgccgcggt gacaccaatt atagaaattt agtccccaaa ctcccaacca ag
//
