LOCUS       sequence-288750-        8599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6Biso1-Nterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8599)
  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 8599)
  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-HsTNRC6Biso1-N-term_F
FEATURES             Location/Qualifiers
     source          1..8599
                     /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             1197..1223
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1203..1229
                     /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    4303..4424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4314..4333)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4368..4387
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4610..5065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4697..4716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4907..4928
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5182..5539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5182..5373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5205..5225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5390..5525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5590..6384
                     /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(5644..5663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6254..6273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6448..6496
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6537..6556)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6656..6678
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6716..6734)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6802..6906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6907..6975,6976..7767)
                     /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(7125..7144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7938..8526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8427..8446
                     /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 aggctgccca agcttacacc atggcctacc
     1201 catatgatgt tccagattac gcttcactcg acgcggatcc cgtcgaattc acgcgtggta
     1261 cctctagagt cgacatgaga gagaaggagc aagaaaggga agaacagtta atggaagaca
     1321 agaaaaggaa gaaagaggat aaaaagaaaa aagaagccac tcagaaggtc acggaacaaa
     1381 aaaccaaagt gcccgaagtg acgaaaccaa gtttaagcca accaacggcc gccagcccaa
     1441 ttggcagctc tccatcgcca ccagtcaatg gtggcaacaa tgccaaaagg gtggcagtgc
     1501 cgaacggaca accgccaagc gccgcccgct acatgcctcg ggaggtgccg ccgcgattcc
     1561 gttgccagca ggaccacaaa gtgttactaa aacgtgggca gccccctcca ccgtcctgca
     1621 tgctccttgg gggtggggca gggcctcctc cctgcacagc acctggagca aacccaaaca
     1681 acgcacaagt gacaggagcg ctgctgcaga gtgagagtgg gactgcgcca gactcaaccc
     1741 ttggaggtgc tgctgcttca aattatgcaa attccacttg gggctcggga gcctcctcca
     1801 acaacggcac ctcccccaac ccaattcaca tctgggacaa ggtgattgta gacgggtctg
     1861 acatggaaga gtggccttgt attgccagca aagacactga atcttcttcc gaaaacacca
     1921 ccgataacaa cagtgcctcg aaccctggct ctgagaagag cactctgcca ggaagcacca
     1981 ctagtaacaa aggaaaaggg agccagtgcc agtctgcaag ttctgggaac gaatgtaatc
     2041 ttggggtctg gaaatctgac cctaaggcta aatctgttca atcttccaac tctactacag
     2101 agaacaacaa tggactagga aattggagga atgtgagtgg tcaggataga attggacctg
     2161 gctctggctt cagcaacttt aacccaaata gcaacccatc tgcctggcca gcactggtcc
     2221 aagaaggaac ttctaggaaa ggggcattgg aaacagataa tagtaattcc agtgcacagg
     2281 ttagcacagt aggtcagaca tccagggaac agcagtcaaa gatggaaaat gcgggtgtta
     2341 attttgttgt ctctggcaga gaacaggctc aaattcataa cactgatgga ccaaaaaatg
     2401 gaaacactaa ctccttgaac ttaagttcac caaaccccat ggagaataag ggaatgccct
     2461 ttggaatggg cttggggaac acctccagga gcactgatgc cccttcacaa agcactggag
     2521 atcgaaagac tgggagtgtt ggatcttggg gtgcagctag ggggccttct ggaactgaca
     2581 cagtctctgg acaaagcaat tctggaaaca atgggaacaa tggaaaagag agagaggact
     2641 cctggaaagg agcttctgtt cagaaatcaa ctgggtcaaa aaatgactct tgggacaaca
     2701 ataacaggtc tacgggtggg tcctggaact ttggccccca ggactctaat gacaacaaat
     2761 ggggtgaagg gaacaaaatg acatctgggg tctctcaggg agaatggaaa cagccgactg
     2821 ggtctgatga gttgaaaatt ggagaatgga gtggtccaaa ccaaccaaat tctagcactg
     2881 gagcatggga caatcaaaag ggccaccccc tccctgaaaa ccaaggcaat gcccaggctc
     2941 cctgttgggg aagatcttcc agctccacag gaagtgaagt tggaggtcaa agcactggaa
     3001 gcaaccacaa agcaggaagt agtgacagtc ataactctgg ccgtcggtcg tacaggccca
     3061 cacatcctga ttgtcaggct gtcttgcaga ctcttttgag ccgaactgat ttggacccca
     3121 gggtgctctc aaacactggc tggggccaaa ctcaaattaa gcaggacaca gtgtgggaca
     3181 ttgaagaggt gccaaggcct gaggggaaat ctgacaaagg aactgagggg tgggagagcg
     3241 ctgccacaca gaccaagaac tcagggggct ggggagatgc acccagccaa agcaatcaaa
     3301 tgaagtctgg atggggggag ctctcagcct ctacagagtg gaaagacccc aagaacacag
     3361 gaggctggaa tgactacaag aacaacaact cttccaactg gggaggagga cgacctgatg
     3421 aaaagacacc ttcctcttgg aatgagaatc ccagcaagga tcaggggtgg ggaggtggac
     3481 gccagcccaa tcaaggatgg tcttctggaa agaatggttg gggggaggaa gtcgatcaga
     3541 caaaaaacag caattgggaa agttctgcaa gtaaacctgt gtctgggtgg ggtgaaggag
     3601 ggcagaatga aatcgggact tggggtaatg gtggcaatgc aagcctagct tcaaaaggtg
     3661 ggtgggagga ttgcaaaaga tccccagcat ggaatgagac gggccgacag cccaattcct
     3721 ggaataaaca acaccaacag cagcagcccc cacagcagcc gccgccacca caaccagagg
     3781 cttctggttc gtggggaggc ccacccccac cacctccagg caacgttcga ccttccaatt
     3841 ccagctggag cagcgggcca cagcctgcaa cacctaagga tgaggaaccc agtggttggg
     3901 aagagccatc cccacagtca attagtcgga aaatggacat tgatgatggc acttcagcat
     3961 ggggagaccc taacagttat aactacaaga atgtgaatct gtgggataag aattcccaag
     4021 ggggcccagc acctcgagaa ccaaacctgc ccaccccaat gaccagtaaa tcggcatcag
     4081 attccaaatc tatgcaagac ggctgggggg agagtgacgg gccagtcaca ggagctcgcc
     4141 atcccagctg ggaagaggag gaggatggag gagtctggaa caccactggc tctcagggca
     4201 gtgcttcctc ccacaactca gcaagctggg gacaaggagg aaagaaacaa atgaaggcgg
     4261 ccgcttccct ttagtgaggg ttaatgcttc gagcagacat gataagatac attgatgagt
     4321 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4381 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4441 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     4501 tctacaaatg tggtaaaatc cgataaggat cgatccgggc tggcgtaata gcgaagaggc
     4561 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt
     4621 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4681 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4741 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4801 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4861 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4921 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4981 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5041 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     5101 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     5161 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     5221 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5281 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5341 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5401 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5461 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5521 aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga acttaaggct
     5581 agagccacca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5641 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5701 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5761 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5821 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5881 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5941 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6001 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6061 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6121 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6181 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6241 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6301 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6361 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6421 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt
     6481 gttggttttt tgtgtgaatc gatagcgata aggatccgcg tatggtgcac tctcagtaca
     6541 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     6601 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     6661 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     6721 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     6781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6841 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6901 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6961 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7021 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7081 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7141 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7201 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7261 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7321 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7381 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7441 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7501 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7561 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7621 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7681 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7741 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7801 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7861 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7921 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7981 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8041 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8101 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8161 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8221 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8281 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8341 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8401 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8461 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8521 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8581 cacatggctc gacagatct
//
