LOCUS       sequence-288962-        6199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6C-DUF-RRM-Cterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6199)
  AUTHORS   Eulalio A, Helms S, Fritzsch C, Fauser M, Izaurralde E
  TITLE     A C-terminal silencing domain in GW182 is essential for miRNA 
            function.
  JOURNAL   RNA. 2009 Jun;15(6):1067-77. doi: 10.1261/rna.1605509. Epub 2009 Apr
            21.
  PUBMED    19383769
REFERENCE   2  (bases 1 to 6199)
  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: 
            "10.1261/rna.1605509"; journalName: "RNA"; date: "2009-06"; volume: 
            "15"; issue: "6"; pages: "1067-77"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsTNRC6C-DUF-RRM-Cterm_H
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1089..1098
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1095..1160
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1179..1205
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1185..1211
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     polyA_signal    1903..2024
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1914..1933)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1968..1987
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2210..2665
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2297..2316)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2507..2528
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2782..3139
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2782..2973
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2805..2825)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2990..3125
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3190..3984
                     /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(3244..3263)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3854..3873
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4048..4096
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4137..4156)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4256..4278
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4316..4334)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4402..4506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4507..4575,4576..5367)
                     /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(4725..4744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5538..6126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6027..6046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag
     1141 ctcaatggaa agctgcaaac ccaccgctcg agatggccta cccatatgat gttccagatt
     1201 acgcttcact cgactctagc atcaactggc ccccagaatt ccatccggga gttccatgga
     1261 aaggactgca gaatattgac cctgagaatg accctgacgt cactcctggc agtagaacca
     1321 gcagctggct cgttcttcga aacctcactc cccagattga tggttctaca ctgcggacat
     1381 tgtgtttgca acatgggcct cttatcacat tccacctgaa tctgactcaa ggcaatgctg
     1441 tggtccggta cagctccaag gaggaggctg ccaaggccca gaagtctctg cacatgtgcg
     1501 tcctgggaaa cactaccatc ctggccgagt tcgctggtga agaagaagtg aatcgcttct
     1561 tagcccaagg ccaggcgctg ccacccactt ccagctggca gtccagcagc gcgtccagcc
     1621 agccgcggct cagcgcagcg ggcagctccc atggcctggt acgcagcgac gctggccact
     1681 ggaacgcccc gtgcctgggt ggcaagggga gcagtgagct gctgtggggc ggggtgcccc
     1741 agtactccag cagcctgtgg ggcccgccca gcgccgacga cagcagggtg ataggcagcc
     1801 ccacgccgct aaccaccctg ctgcctgggg acctgctcag cggggagtcc ctgtaggcgg
     1861 ccgcttccct ttagtgaggg ttaatgcttc gagcagacat gataagatac attgatgagt
     1921 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1981 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2041 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     2101 tctacaaatg tggtaaaatc cgataaggat cgatccgggc tggcgtaata gcgaagaggc
     2161 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt
     2221 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2281 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2341 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2401 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2461 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2521 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2581 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2641 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     2701 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     2761 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     2821 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2881 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2941 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3001 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3061 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3121 aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga acttaaggct
     3181 agagccacca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3241 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3301 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3361 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3421 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3481 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3541 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3601 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3661 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3721 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3781 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3841 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3901 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3961 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4021 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt
     4081 gttggttttt tgtgtgaatc gatagcgata aggatccgcg tatggtgcac tctcagtaca
     4141 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4201 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4261 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4321 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4381 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4441 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4501 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4561 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4621 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4681 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4741 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4801 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4861 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4921 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4981 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5041 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5101 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5161 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5221 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5281 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5341 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5401 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6181 cacatggctc gacagatct
//
