LOCUS       sequence-453675-        8542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6Biso3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8542)
  AUTHORS   Lazzaretti D, Tournier I, Izaurralde E
  TITLE     The C-terminal domains of human TNRC6A, TNRC6B, and TNRC6C silence 
            bound transcripts independently of Argonaute proteins.
  JOURNAL   RNA. 2009 Jun . 15(6):1059-66.
  PUBMED    19383768
REFERENCE   2  (bases 1 to 8542)
  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: "RNA. 2009 
            Jun . 15(6):1059-66."
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsTNRC6Biso3_C
FEATURES             Location/Qualifiers
     source          1..8542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      952..961
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             958..1023
                     /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             1060..1086
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1066..1092
                     /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    4109..4230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4120..4139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4174..4193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4416..4871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4503..4522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4713..4734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4988..5345
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4988..5179
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5011..5031)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5196..5331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5396..6190
                     /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(5450..5469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6060..6079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6254..6302
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6343..6362)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6462..6484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6522..6540)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6608..6712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6713..6781,6782..7573)
                     /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(6931..6950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7744..8332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8233..8252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agccaccatg
      961 gacgcacaaa cacgacgacg tgagcgtcgc gctgagaaac aagctcaatg gaaagctgca
     1021 aacccaccgc tcgaggctgc ccaagcttac accatggcct acccatatga tgttccagat
     1081 tacgcttcac tcgacgcgga tcccgtcgaa ttcacgcgtg gtacctctag agtcgacacc
     1141 aatgagggag aagtatcgga agaaagcagt tccaaggtgg aacaggagga ttttgtaatg
     1201 gaagggcatg gcaagactcc acctcctggt gaagaaagca aacaagagaa ggagcaagaa
     1261 agggaagaac agttaatgga agacaagaaa aggaagaaag aggataaaaa gaaaaaagaa
     1321 gccactcaga aggtcacgga acaaaaaacc aaagtgcccg aagtgacgaa accaagttta
     1381 agccaaccaa cggccgccag cccaattggc agctctccat cgccaccagt caatggtggc
     1441 aacaatgcca aaagggtggc agtgccgaac ggacaaccgc caagcgccgc ccgctacatg
     1501 cctcgggagg tgccgccgcg attccgttgc cagcaggacc acaaagtgtt actaaaacgt
     1561 gggcagcccc ctccaccgtc ctgcatgctc cttgggggtg gggcagggcc tcctccctgc
     1621 acagcacctg gagcaaaccc aaacaacgca caagtgacag gagcgctgct gcagagtgag
     1681 agtgggactg cgccagattc caaatctatg caagacggct ggggggagag tgacgggcca
     1741 gtcacaggag ctcgccatcc cagctgggaa gaggaggagg atggaggagt ctggaacacc
     1801 actggctctc agggcagtgc ttcctcccac aactcagcaa gctggggaca aggaggaaag
     1861 aaacaaatga agtgctcact caaaggagga aacaatgatt catggatgaa tcctcttgcc
     1921 aaacagtttt caaatatggg attgctgagt cagactgaag ataatccaag cagcaaaatg
     1981 gatttgtctg taggaagcct ttcagataaa aaatttgatg tggacaagcg agcgatgaat
     2041 ctcggggatt ttaatgatat catgaggaag gatcgatctg ggttccgtcc acctaattcc
     2101 aaagacatgg gaaccacaga tagtgggcct tattttgaga agggcggtag tcatggtttg
     2161 tttggaaaca gcacagcaca atcgagaggt ctgcacacac ccgtgcagcc actaaattct
     2221 tctcccagtc tccgggcgca agtgcctccc cagtttattt ccccccaggt ttctgcctca
     2281 atgctcaagc agtttcccaa cagtggcctg agtccaggtc ttttcaatgt ggggccccag
     2341 ttatctcctc aacaaattgc catgctgagc cagcttccac aaattcccca gtttcagttg
     2401 gcatgtcagc ttctcttgca gcagcagcaa cagcagcagt tgttacagaa ccagagaaag
     2461 atttctcaag ctgtacgcca acagcaagag cagcagctgg ctcgaatggt gagtgcactg
     2521 cagcagcagc agcagcagca gcagaggcag ccaggcatga agcactcgcc ctctcatcct
     2581 gttgggccca agccgcatct ggacaacatg gtacccaacg cattgaatgt ggggctccca
     2641 gaccttcaaa ccaaagggcc aatacctgga tatggttctg gcttcagctc tggcggcatg
     2701 gactatggca tggttggtgg gaaggaggct ggaaccgagt ctcgctttaa acagtggacc
     2761 tccatgatgg aggggctgcc ctctgtagcc acacaggaag ccaatatgca caaaaatggc
     2821 gctatagtgg cccctggtaa aacccgggga gggtcaccgt acaaccagtt tgatatcatc
     2881 cctggtgaca cactgggtgg ccatacgggt cctgctggtg atagctggtt acctgccaaa
     2941 tctccaccaa caaataaaat cggaagtaaa tccagcaatg ccagttggcc tccagaattc
     3001 caaccaggag tgccatggaa aggtatccaa aacattgacc ctgaatctga cccctatgtc
     3061 accccaggaa gtgtgctggg gggtacagcc acatctccca ttgtagatac tgaccaccaa
     3121 ctgctgcggg ataacaccac agggtctaat tcttccctca acacctcgct gccttcacct
     3181 ggtgcctggc cctacagtgc ctctgacaac tcctttacca acgttcatag cacttcagca
     3241 aagttccctg attacaaatc aacatggtcc ccagatccca taggacacaa ccccactcat
     3301 ctctccaaca agatgtggaa aaaccatatt tcctccagga acactacacc gctgccccgc
     3361 ccacctcctg gtctgaccaa ccccaaacca tcatctccct ggagcagcac agcaccccga
     3421 tcagtcaggg ggtgggggac acaggactca cggctcgcct cggcctctac ctggagtgat
     3481 ggtggctcag ttcgtcctag ttactggctg gttcttcaca atctcacccc acagattgat
     3541 gggtcaacct tgagaacgat ctgcatgcag catggcccac tgctgacatt ccatctgaat
     3601 ctaacccagg gcactgccct gatccgatac agcaccaaac aggaggcggc caaggcccaa
     3661 actgcactgc acatgtgtgt gttgggaaac actaccatcc ttgctgagtt tgccactgat
     3721 gatgaagtca gccgctttct ggcacaagct cagcccccta cacctgcagc aaccccaagt
     3781 gcgccagctg cggggtggca gtcgctggag accggccaga accagtcaga tcccgtggga
     3841 cctgctctga atctttttgg tgggtccact gggctcgggc agtggagcag cagcgctggt
     3901 ggcagcagcg gggccgatct tgctggcgct tcattgtggg ggcccccaaa ctattcttct
     3961 agcttatggg gagtcccaac ggtggaagat ccccatagga tgggcagccc tgctccttta
     4021 ctacctggtg accttctggg aggagggtcg gattcaatct gagcggccgc ttccctttag
     4081 tgagggttaa tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     4141 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4201 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4261 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4321 aaaatccgat aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc
     4381 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
     4441 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4501 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4561 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4621 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4681 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4741 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4801 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4861 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4921 atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     4981 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5041 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5101 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5161 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5221 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5281 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5341 gcaaaaagct tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat
     5401 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5461 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5521 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     5581 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5641 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5701 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5761 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5821 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5881 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     5941 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6001 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6061 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6121 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6181 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6241 tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     6301 tgaatcgata gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc
     6361 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6421 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6481 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     6541 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     6601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6781 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7561 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7621 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7681 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7741 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7801 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7861 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7921 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7981 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     8041 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     8101 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     8161 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8221 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8281 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8341 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca
     8401 gatcttcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt
     8461 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     8521 tgtccaatat gaccgccatg tt
//
