LOCUS       sequence-356082-        6055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing various parts of TRBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6055)
  AUTHORS   Takahashi T, Nakano Y, Onomoto K, Murakami F, Komori C, Suzuki Y, 
            Yoneyama M, Ui-Tei K
  TITLE     LGP2 virus sensor regulates gene expression network mediated by 
            TRBP-bound microRNAs.
  JOURNAL   Nucleic Acids Res. 2018 Sep 28;46(17):9134-9147. doi: 
            10.1093/nar/gky575.
  PUBMED    29939295
REFERENCE   2  (bases 1 to 6055)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2018-09-28- 28"; volume: "46"; issue: 
            "17"; pages: "9134-9147"
COMMENT     Sequence Label: pcDNA3.1-TRBP-L2+D2+L3-myc/His
FEATURES             Location/Qualifiers
     source          1..6055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        108..487
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        488..691
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     642..662
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     736..755
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        736..754
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1432..1461
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1477..1494
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1517..1534)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1523..1747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1793..2221
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1880..1899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2090..2111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2230..2250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2235..2564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2415..2550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2477..2496
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2631..3425
                     /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(2685..2704)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3295..3314
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3601..3722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3638..3657)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3692..3711
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3771..3787)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3771..3787)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3784..3806)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3795..3811
                     /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(3819..3825,3826..3843,3844..3849))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3864..3885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4002..4019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4173..4761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4253..4272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4932..5723,5724..5792))
                     /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     5555..5574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5793..5897)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5972..5991)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
       61 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      121 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      181 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      241 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      301 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      361 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      421 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      481 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      541 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      601 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      661 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      721 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agttaagctt
      781 atggggagca tgctggagcc ggccctggag gacagcagtt ctttttctcc cctagactct
      841 tcactgcctg aggacattcc ggtttttact gctgcagcag ctgctacccc agttccatct
      901 gtagtcctaa ccaggagccc ccccatggaa ctgcagcccc ctgtctcccc tcagcagtct
      961 gagtgcaacc ccgttggtgc tctgcaggag ctggtggtgc agaaaggctg gcggttgccg
     1021 gagtacacag tgacccagga gtctgggcca gcccaccgca aagaattcac catgacctgt
     1081 cgagtggagc gtttcattga gattgggagt ggcacttcca aaaaattggc aaagcggaat
     1141 gcggcggcca aaatgctgct tcgagtgcac acggtgcctc tggatgcccg ggatggcaat
     1201 gaggtggagc ctgatgatga ccacttctcc attggtgtgg gctcccgcct ggatggtctt
     1261 cgaaaccggg gcccaggttg cacctgggat tctctacgaa attcagtagg agagaagatc
     1321 ctgtccctcc gcagttgctc cctgggctcc ctgggtgccc tgggccctgc ctgctgccgt
     1381 gtcgcggccg ctgcaagctc ctcgagcatg catctagagg gcccgcggtt cgaacaaaaa
     1441 ctcatctcag aagaggatct gaatatgcat accggtcatc atcaccatca ccattgagtt
     1501 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1561 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1621 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1681 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1741 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     1801 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1861 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1921 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1981 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2041 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2101 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2161 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2221 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2281 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2341 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2401 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2461 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     2521 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     2581 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     2641 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2701 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2761 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2821 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2881 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2941 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3001 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3061 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3121 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3181 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3241 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3301 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3361 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3421 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     3481 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3541 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     3601 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3661 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3721 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     3781 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3841 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3901 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3961 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4021 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4081 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4141 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4201 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4261 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4321 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4381 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4441 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4501 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4561 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4621 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4681 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4741 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4801 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4861 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4921 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4981 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5041 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5101 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5161 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5221 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5281 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5341 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5401 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5461 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5521 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5581 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5641 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5701 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5761 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5881 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     5941 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6001 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaa
//
