LOCUS       sequence-288961-        5929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6C-DUF-RRM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5929)
  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 5929)
  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_H
FEATURES             Location/Qualifiers
     source          1..5929
                     /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    1633..1754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1644..1663)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1698..1717
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1940..2395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2027..2046)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2237..2258
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2512..2869
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2512..2703
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2535..2555)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2720..2855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2920..3714
                     /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(2974..2993)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3584..3603
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3778..3826
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3867..3886)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3986..4008
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4046..4064)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4132..4236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4237..4305,4306..5097)
                     /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(4455..4474)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5268..5856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5757..5776
                     /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 ccataggcgg ccgcttccct ttagtgaggg ttaatgcttc
     1621 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1681 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1741 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     1801 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc cgataaggat
     1861 cgatccgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     1921 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1981 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2041 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2101 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2161 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2221 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2281 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2341 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     2401 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     2461 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     2521 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2581 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2641 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2701 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2761 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2821 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgattc
     2881 ttctgacaca acagtctcga acttaaggct agagccacca tgattgaaca agatggattg
     2941 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3001 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3061 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3121 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3181 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3241 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3301 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3361 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3421 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3481 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3541 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3601 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3661 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3721 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat
     3781 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata
     3841 aggatccgcg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3901 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3961 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4021 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4081 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4141 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4201 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4261 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4321 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4381 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4441 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4501 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4561 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4621 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4681 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4741 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4801 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4861 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4921 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4981 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5041 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5101 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5161 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5221 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5281 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5341 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5401 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5461 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5521 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5581 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5641 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5701 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5761 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5821 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5881 cggttcctgg ccttttgctg gccttttgct cacatggctc gacagatct
//
