LOCUS       sequence-288953-        9288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6A_1-1248.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9288)
  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 9288)
  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-HsTNRC6A_1-1248_H
FEATURES             Location/Qualifiers
     source          1..9288
                     /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"
     primer_bind     complement(5007..5026)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5061..5080
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5299..5754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5386..5405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5596..5617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5871..6228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5871..6062
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5894..5914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6079..6214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6279..7073
                     /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(6333..6352)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6943..6962
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7137..7185
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(7226..7245)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7345..7367
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7405..7423)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7491..7595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7596..7664,7665..8456)
                     /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(7814..7833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8627..9215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9116..9135
                     /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 agatggctta cccatacgat gttccagatt
     1201 acgctatgga tgctgattct gcctccagtt ctgaatcaga gagaaacatc actatcatgg
     1261 cttcagggaa cacaggtggt gaaaaagatg gccttcggaa tagcactgga cttggttccc
     1321 aaaacaagtt tgtagttggt agcagcagca ataatgtggg ccatggaagt agtactgggc
     1381 catggggttt ttcccatgga gccataataa gcacatgtca ggtctctgtg gatgctcctg
     1441 aaagcaaatc tgaaagtagc aacaatagaa tgaatgcttg gggcactgta agttcttcat
     1501 caaatggagg gttaaatcca agcactttga attcagctag caaccatggt gcctggccag
     1561 tattagagaa caatggactt gccctaaaag ggcctgtagg gagtggtagt tctggcatta
     1621 atattcagtg cagtactata ggccagatgc ctaacaatca gagtattaac tctaaagtga
     1681 gtggtggttc tacccatggt acctggggaa gccttcagga aacttgtgaa tctgaagtaa
     1741 gtggtacaca gaaggtttca ttcagtggtc aacctcaaaa tattaccact gaaatgactg
     1801 gaccaaataa cactactaac tttatgacct ctagtttacc aaactccggt tcagtgcaga
     1861 ataatgagct gcctagtagt aacacagggg cctggcgtgt gagcacaatg aatcatcctc
     1921 agatgcaggc tccatcaggt atgaatggca cttccctttc tcaccttagc aatggagagt
     1981 caaaaagtgg aggctcttat ggtactacat ggggtgccta tggttctaat tactctggag
     2041 acaaatgttc aggccctaat ggccaagcta atggtgacac tgtgaatgca actctaatgc
     2101 agcctggcgt aaatggtcct atgggcacta actttcaagt taacacaaac aaaggaggtg
     2161 gtgtgtggga atctggtgca gcaaactccc agagtacatc atggggaagt ggaaatggcg
     2221 caaattctgg aggaagtcga agaggatggg gaacccctgc acaaaacact ggcactaatt
     2281 tacccagcgt tgagtggaac aaactgccta gcaatcagca ttccaatgat agtgcaaatg
     2341 gcaatggtaa gacgtttaca aatggatgga aatctactga ggaagaggat cagggttctg
     2401 ccacatctca gacaaatgag caaagcagtg tgtgggccaa aacaggaggt acagtggaga
     2461 gcgatggtag tacagaaagc actggacgcc ttgaggaaaa aggaactggg gaaagtcaga
     2521 gtagagacag aagaaaaatt gatcagcaca cattactcca aagcattgta aacagaactg
     2581 acttagatcc acgtgtcctg tccaactctg gttggggaca gactcctatt aagcagaata
     2641 ctgcctggga tacagaaaca tcacctagag gggaacgaaa gactgacaat gggacagagg
     2701 cctggggaag ctctgcaaca cagactttta actcaggggc atgtatagat aagactagcc
     2761 ctaatggtaa tgatacctca tctgtatcag ggtggggcga tcccaaacct gctctgaggt
     2821 ggggagattc caaaggctca aactgccagg gggggtggga agatgattct gctgctacag
     2881 gaatggtcaa gagcaatcag tgggggaatt gcaaagagga gaaggctgca tggaatgact
     2941 cgcaaaagaa taaacaggga tggggtgatg gacaaaaatc aagccaaggg tggtctgttt
     3001 ctgccagtga taactgggga gaaacttcaa ggaataacca ttggggtgag gccaataaga
     3061 aatccagctc aggaggtagt gacagtgaca ggtccgtttc cggttggaac gaacttggta
     3121 aaactagttc tttcacttgg ggaaacaaca taaatccaaa taattcatca ggatgggatg
     3181 aatcttctaa acctactcct tcccagggat ggggagaccc tccaaagtct aatcagtctc
     3241 taggttgggg agattcgtca aagccagtca gctctccaga ctggaacaag caacaagaca
     3301 ttgttggatc ttggggaatc ccaccagcta caggcaaacc tcctggtaca ggctggctgg
     3361 ggggacctat accagcccca gcaaaagaag aagaacccac aggctgggag gaaccatccc
     3421 cagaatctat acgtcgcaaa atggagattg atgatggaac ttcagcttgg ggagatccaa
     3481 gcaaatacaa ctacaaaaat gtgaacatgt ggaacaaaaa cgtcccaaat ggcaacagcc
     3541 gttcagacca gcaagcacag gtacatcagc tgctaacgcc tgcaagtgcc atctcaaaca
     3601 aagaggcaag cagtggctct ggctggggtg agccctgggg ggagccttct actccagcca
     3661 caactgtgga taatggtact tcagcatggg gtaagcccat agacagtggt cccagctggg
     3721 gggaacccat tgctgcggca tccagcacat ccacgtgggg ctccagctct gttggtccac
     3781 aagcattaag caaatctggg ccaaaatcta tgcaagatgg ctggtgtggt gatgatatgc
     3841 cattgcctgg aaatcgcccc actggctggg aagaggaaga ggatgtggag attggaatgt
     3901 ggaatagtaa ttcatctcaa gagcttaact catctttaaa ttggccacca tatacaaaga
     3961 aaatgtcatc gaagggtctg agtggcaaaa aaaggagaag ggaaagggga atgatgaaag
     4021 gtggaaacaa acaagaagaa gcgtggataa atccatttgt taaacagttt tcaaacatca
     4081 gtttttcgag agactcacca gaggaaaatg tacaaagcaa taagatggac ctttctggag
     4141 gaatgttaca agacaaacga atggagatag ataaacatag cctaaatatt ggtgattaca
     4201 atcgaacggt cgggaaaggc cctggttctc ggcctcagat ttccaaagag tcttccatgg
     4261 agcgcaatcc ttattttgat aaggatggca ttgtagcaga tgaatcccaa aacatgcagt
     4321 ttatgtccag tcaaagcatg aagcttcccc cttcaaatag tgcactacct aaccaggccc
     4381 ttggctccat agcagggctg ggtatgcaaa acttgaattc tgttagacag aatggcaatc
     4441 ccagtatgtt tggtgttgga aacacagcag cacaaccccg gggcatgcag cagcctccag
     4501 cacaacctct tagttcatct cagcctaatc tccgtgctca agtgcctcct ccattactct
     4561 cccctcaggt tccagtttca ttgctgaagt atgcaccaaa caacggtggc ctgaatccac
     4621 tctttggccc tcaacaggta gccatgctga accagctatc ccagctaaac cagctttctc
     4681 agatctccca gttacagcga ttgttagcgc agcagcaaag ggcgcagagt cagagaagcg
     4741 tgccttctgg gaaccggccg cagcaagacc agcagggtcg acctcttagt gtgcagcagc
     4801 aaatgatgca acaatctcgt caacttgatc caaacctgtt ggtgaagcag cagactccac
     4861 catctcagca gcagccactc catcagccag ccatgaagtc tttccttgac aatgtcatgc
     4921 cccacactac acctgagctg caaaaagggg cggccgcttc cctttagtga gggttaatgc
     4981 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     5041 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     5101 gctgcaataa acaagtaaca acaattgcat tcattttatg tttcaggttc agggggagat
     5161 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc
     5221 gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5281 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5341 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5401 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5461 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5521 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5581 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5641 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5701 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     5761 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     5821 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     5881 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6061 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6121 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6181 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct
     6241 tctgacacaa cagtctcgaa cttaaggcta gagccaccat gattgaacaa gatggattgc
     6301 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6361 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6421 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     6481 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6541 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6601 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6661 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6721 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6781 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     6841 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6901 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6961 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     7021 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     7081 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata
     7141 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa
     7201 ggatccgcgt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     7261 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     7321 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     7381 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     7441 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     7501 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     7561 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     7621 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     7681 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     7741 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7801 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7861 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7921 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7981 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     8041 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     8101 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     8161 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     8221 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     8281 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     8341 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     8401 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     8461 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     8521 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     8581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     8641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     8701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8761 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     9001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     9061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     9121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     9181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     9241 ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatct
//
