LOCUS       sequence-438204-        6333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TRBP-V336A/H338A.
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-TRBP-V336A/H338A
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      669..1257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1158..1177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1411..1428
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1545..1566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1581..1586,1587..1604,1605..1611)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1619..1635
                     /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)."
     primer_bind     1624..1646
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1643..1659
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1643..1659
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1708..1829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1719..1738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1773..1792
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1959..2981)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(1959..2981)
                     /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
                     "
     protein_bind    complement(2989..3036)
                     /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)."
     primer_bind     3168..3187
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3320..3544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3533..3550
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     3577..3596
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3577..3595
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3651..3667
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3652..3668
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4784..4810)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4811..4825)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     regulatory      4837..4846
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4942..4966)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    4976..4994
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4997..5015
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(5017..5220)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5046..5066)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5221..5600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5772..5791
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5866..5970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5971..6039,6040..6333,1..498)
                     /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     complement(6189..6208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
       61 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      121 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      181 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      241 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      301 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      361 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      421 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      481 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      601 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      661 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      721 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      781 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1141 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1201 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1261 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1321 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1381 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1441 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     1501 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     1561 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     1621 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     1681 ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa
     1741 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1801 taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc
     1861 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     1921 tagaaggcgg cggtggaatc gaaatctcgt agtacgtgct attcctttgc cctcggacga
     1981 gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg
     2041 gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt
     2101 gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga
     2161 tagagttggt caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc
     2221 tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc
     2281 cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca
     2341 atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc
     2401 acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc
     2461 gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca
     2521 gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat
     2581 gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat ggcctccgcg
     2641 accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt
     2701 gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc
     2761 ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca
     2821 tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca
     2881 cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc
     2941 agaaacttct cgacagacgt cgcggtgagt tcaggctttt tggccaagga agttcctata
     3001 ctttctagag aataggaact tcggaatagg aacttctagg tacgtgaacc atcaccctaa
     3061 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3121 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3181 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3241 cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat accccctaga gccccagctg
     3301 gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct
     3361 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     3421 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     3481 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     3541 acagtcgagg ctgatcagcg ggtttcgagt gaattgtaat acgactcact atagggcgaa
     3601 ttgggtacac ttacctggta ccttaattaa ttaaggtacc gggccccccc tcgaggtcga
     3661 cggtatcgat aagctttcac ttgctgcctg ccatgatctt gaggtactgc agggcacggc
     3721 gggcagcctc accacgggct gcctccctgg tggttgcaga gcctgcacat gcagtggccg
     3781 gctgggtgga cagttccacc aggcactggc agagtccact caggctcagc tcctcaatat
     3841 ccaggtagct gacgtgaaag gcctgctcct cagagagctc actgaggaca cggcagcagg
     3901 cagggcccag ggcacccagg gagcccaggg agcaactgcg gagggacagg atcttctctc
     3961 ctactgaatt tcgtagagaa tcccaggtgc aacctgggcc ccggtttcga agaccatcca
     4021 ggcgggagcc cacaccaatg gagaagtggt catcatcagg ctccacctca ttgccatccc
     4081 gggcatccag aggcaccgtg tgcactcgaa gcagcatttt ggccgccgca ttccgctttg
     4141 ccaatttttt ggaagtgcca ctcccaatct caatgaaacg ctccactcga caggtcatgg
     4201 tgaattcttt gcggtgggct ggcccagact cctgggtcac tgtgtactcc ggcaaccgcc
     4261 agcctttctg caccaccagc tcctgcagag caccaacggg gttgcactca gactgctgag
     4321 gggagacagg gggctgcagt tccatggggg ggctcctggt taggactaca gatggaactg
     4381 gggtagcagc tgctgcagca gtaaaaaccg gaatgtcctc aggcagtgaa gagtctaggg
     4441 gagaaaaaga actgctgtcc tccagggccg gctccagcat gctcccccct ttgaggtgtt
     4501 tgagggccac ctcagctgcc ttgtgcttgg ctgccttctt gctggggccc tgaccagtgc
     4561 agctggtgtc gccaacggtg acccggaagg tgaaattagg ctggtgggct tggccctcgg
     4621 ctttgagaag gtcgtacaca ggcgtcttcc ctattctggt cccatactcc tgcagaaggc
     4681 tgatcggggt cttgcctggg ttggcggcca gcatttgctc tatactaggc agcccgcagc
     4741 ccgtggtagt gccggagcct tgctcctctt cactcatgct agcagcgtaa tctggaacat
     4801 cgtatgggta cttatcgtcg tcgtcatcct tgtaatccat ggtggcggcc gccaccgcgg
     4861 tggaaacgct agagtccgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca
     4921 gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctcgtc gacgatctct
     4981 atcactgata gggagatctc tatcactgat agggagagct ctgcttatat agacctccca
     5041 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     5101 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     5161 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     5221 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     5281 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     5341 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     5401 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     5461 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     5521 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     5581 actagtcaat aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa
     5641 acgcctaacc ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc
     5701 ttaaattttg ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg
     5761 gcttaactat gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga
     5821 tctcccgatc cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5881 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5941 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6001 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6061 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6121 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6181 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6241 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6301 cttacggatg gcatgacagt aagagaatta tgc
//
