LOCUS       sequence-438208-        6333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TRBP-L326A/Y358A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  AUTHORS   Komori C, Takahashi T, Nakano Y, Ui-Tei K
  TITLE     TRBP-Dicer interaction may enhance HIV-1 TAR RNA translation via TAR
            RNA processing, repressing host-cell apoptosis.
  JOURNAL   Biol Open. 2020 Feb 25;9(2):bio050435. doi: 10.1242/bio.050435.
  PUBMED    32051109
REFERENCE   2  (bases 1 to 6333)
  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: "Biol 
            Open."; date: "2020-02-25"; volume: "9(2):bio050435. doi"; pages: " 
            10.1242/bio.050435"
COMMENT     Sequence Label: pcDNA5-Flag/HA-TRBP-L326A/Y358A
FEATURES             Location/Qualifiers
     source          1..6333
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      989..998
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1010..1024
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             1025..1051
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2167..2183)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2168..2184)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2239..2258)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2240..2258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2285..2302)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2291..2515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2648..2667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2799..2846
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2854..3876
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2854..3876
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4006..4127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4043..4062)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4097..4116
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4176..4192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4176..4192)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4189..4211)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4200..4216
                     /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(4224..4230,4231..4248,4249..4254))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4269..4290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4407..4424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4578..5166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4658..4677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5337..6128,6129..6197))
                     /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     5960..5979
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6198..6302)
                     /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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt tccaccgcgg tggcggccgc caccatggat tacaaggatg acgacgacga
     1021 taagtaccca tacgatgttc cagattacgc tgctagcatg agtgaagagg agcaaggctc
     1081 cggcactacc acgggctgcg ggctgcctag tatagagcaa atgctggccg ccaacccagg
     1141 caagaccccg atcagccttc tgcaggagta tgggaccaga atagggaaga cgcctgtgta
     1201 cgaccttctc aaagccgagg gccaagccca ccagcctaat ttcaccttcc gggtcaccgt
     1261 tggcgacacc agctgcactg gtcagggccc cagcaagaag gcagccaagc acaaggcagc
     1321 tgaggtggcc ctcaaacacc tcaaaggggg gagcatgctg gagccggccc tggaggacag
     1381 cagttctttt tctcccctag actcttcact gcctgaggac attccggttt ttactgctgc
     1441 agcagctgct accccagttc catctgtagt cctaaccagg agccccccca tggaactgca
     1501 gccccctgtc tcccctcagc agtctgagtg caaccccgtt ggtgctctgc aggagctggt
     1561 ggtgcagaaa ggctggcggt tgccggagta cacagtgacc caggagtctg ggccagccca
     1621 ccgcaaagaa ttcaccatga cctgtcgagt ggagcgtttc attgagattg ggagtggcac
     1681 ttccaaaaaa ttggcaaagc ggaatgcggc ggccaaaatg ctgcttcgag tgcacacggt
     1741 gcctctggat gcccgggatg gcaatgaggt ggagcctgat gatgaccact tctccattgg
     1801 tgtgggctcc cgcctggatg gtcttcgaaa ccggggccca ggttgcacct gggattctct
     1861 acgaaattca gtaggagaga agatcctgtc cctccgcagt tgctccctgg gctccctggg
     1921 tgccctgggc cctgcctgct gccgtgtcct cagtgagctc tctgaggagc aggcctttca
     1981 cgtcagctac ctggatattg aggagctgag cctgagtgga ctctgccagt gcgcagtgga
     2041 actgtccacc cagccggcca ctgtgtgtca tggctctgca accaccaggg aggcagcccg
     2101 tggtgaggct gcccgccgtg ccctgcaggc actcaagatc atggcaggca gcaagtgaaa
     2161 gcttatcgat accgtcgacc tcgagggggg gcccggtacc ttaattaatt aaggtaccag
     2221 gtaagtgtac ccaattcgcc ctatagtgag tcgtattaca attcactcga aacccgctga
     2281 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2341 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2401 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2461 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2521 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2581 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2641 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2701 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2761 cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt ccgaagttcc
     2821 tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac cgcgacgtct
     2881 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     2941 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     3001 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     3061 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     3121 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     3181 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     3241 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     3301 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     3361 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     3421 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     3481 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     3541 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     3601 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     3661 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     3721 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     3781 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     3841 cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga tttcgattcc
     3901 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     3961 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4021 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4081 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4141 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4201 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4261 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4321 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4381 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4441 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4501 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4561 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4621 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4681 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4741 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4801 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4861 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4921 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4981 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5041 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5101 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5161 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5221 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5281 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5341 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5401 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5461 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5521 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5581 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5641 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5701 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5761 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5821 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5881 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5941 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6001 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6061 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6121 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6181 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6241 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6301 cgcacatttc cccgaaaagt gccacctgac gtc
//
