LOCUS       sequence-351336-        7542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KSHV Downstream Mir-SYN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7542)
  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 7542)
  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: pOME0133
FEATURES             Location/Qualifiers
     source          1..7542
                     /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"
     promoter        complement(3095..3113)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3096..3113)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3133..3150)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3139..3363
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3409..3837
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3496..3515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3706..3727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3846..3866)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3851..4180
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4031..4166
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4093..4112
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4247..5041
                     /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(4301..4320)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4911..4930
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5215..5336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5252..5271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5306..5325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5385..5401)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5385..5401)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5398..5420)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5409..5425
                     /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(5433..5439,5440..5457,5458..5463))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5478..5499
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5616..5633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5787..6375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5867..5886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6546..7337,7338..7406))
                     /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     7169..7188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7407..7511)
                     /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 ccgccaaacg ggcgatggaa acttgtcaac actccaaaaa aatagaaggt
      961 tgggtaactg cgggctggtg tcaacttctg ttaattatga atagtgtttt taaagcgggc
     1021 gttcgtaagc gtctggatcg acacaaaccc tgcgcgggtt tagaaagact tgtccagcag
     1081 cacctaatcc atcggcggtg cgggctgatg gttttcgggc tgttgagcga gtctttttat
     1141 ctagtcgccc gttattgttc tgtgtgttct tcatcatttc acccaccgtc tctcaatagc
     1201 cttgggaatc ctacctccac gctcgcgtat gcctcatgcg aggttcgctt tggtcacagc
     1261 ttaaacattt ctagggcggt gttatgatcc ttaatgctta gcctgtgtcc gatgcggccc
     1321 cgtgccttgc ggaagcgcgg gcgggttccc gaccttcacg gcggcaattt ttgtcgtatg
     1381 cgcctcccct cgcctacaag tccaataccc atgagaccac cgtacgcgtg cccaccgatg
     1441 agataccacg cagccgcgca tattggcgtt gagcgccacc ggacggggat ttatgctgta
     1501 tcttactacc atgatcccat gttgctggcg ctcacggccc gtgtgccagc cgcctggacg
     1561 tcgaaagcac gcaaagctga acgttcgggc ttagtgtagc gcgaaccata cgaccttggt
     1621 cccctggcca aagaggggcg caaaacaatt gtgggtggct atcctgccca gagtcccata
     1681 tattctggtg ggtagctaac ctgcccagga cgtccaaata tgccaaatgg taccttagtg
     1741 tctactttgc atatttggaa cccgtgtatc catgtatggt tatcgtgccc agggtgccca
     1801 aattgctcaa taatgaaaat tccattctag cgctggggtg acaaagcatg cactggaaat
     1861 cggccaagtc agcgcagcgc gcgcactccc tcactaacgc cccgcttttg tctgttggaa
     1921 gcagctaggc gcgactgaga gagcacgcgc gcggcgccca cgccggcgcc tggcacgcgg
     1981 ccaacataaa gtgtggatgg ccctgctatc tgactaattt gcatgcgcta atccttttat
     2041 gtgcataaat tatgtagatg aggagtcgcg catgcgcaga aaaattcaga gcgcccgggt
     2101 gcacggggtc acctccaggt cacgccgcta ggtgggaccg tgagcgactc gaaaaattat
     2161 aatttttggc catttcatgg gcgccgccat cttgaatttg ctaatccccc ataatcctct
     2221 gccccgctcc cattggtccg ccggcccgtc aatcaaagtt ttccgagccg ccattggccc
     2281 attcggccga ccaatctcgt tcgagctagg cgaccgcgcc attccattgg acgccccagc
     2341 cgtcaatcaa attcggaggc ctcccattgg cccctatccc tagaactccc aagctgattg
     2401 gcccacagcg ggaaccaatc agcgattaga gttttgtttt gatttttcct atatatatat
     2461 atataatcct ttaatcctag cgcagctgag tcatcgcagc ccctattcca gtaggtatac
     2521 ccagctgggt ctacccagct gcgtaaaccc cgctgcgtaa acacagctgg gtatacgcag
     2581 ctgcgtaaac ccggctgggt aaatccagct gtaattctac aattaaacaa ggtataaacc
     2641 aatggggtac tcaggtggta tttccggaag cattaccaaa taaggcaacc tcagctggga
     2701 ataccagcgg actaccccca actgtattca accctccttc gttttccgga agtatatcca
     2761 tttatggaaa ccagctgggt cactctactg ggatattcta tataataggg cctgatgagt
     2821 catggggttg ggatttttct actgggtcgt ttcggtagat gggggccagg attatagggg
     2881 ccccgcccac cgccactcca gccgtgttag ccaagcgtgg ttgttcgatg ggaggggagc
     2941 tagtgagtca cggccctgga atctcgcctc tgggtggttt cggtaaatgg gagcgggatg
     3001 ctaggtccac gctcacctct ggcgcggccc cggggaattc tgcagatatc catcacactg
     3061 gcggccgctc gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag
     3121 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3181 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3241 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3301 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3361 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3421 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3481 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3541 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3601 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3661 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3721 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3781 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3841 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3901 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3961 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4021 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4081 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4141 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4201 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4261 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4321 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4381 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4441 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4501 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4561 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4621 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4681 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4741 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4801 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4861 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4921 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4981 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5041 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5101 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5161 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5221 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5281 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5341 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5401 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5461 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5521 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5581 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5641 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5701 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5761 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5821 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5881 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5941 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6001 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6061 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6121 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6181 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6241 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6301 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6361 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6421 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6481 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6541 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6601 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6661 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6721 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6781 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6841 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6901 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6961 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7021 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7081 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7141 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7201 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7261 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7321 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7381 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7441 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7501 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
