LOCUS       sequence-288745-        8412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6A-Nterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8412)
  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 8412)
  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-HsTNRC6A-N-term_F
FEATURES             Location/Qualifiers
     source          1..8412
                     /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    4116..4237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4127..4146)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4181..4200
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4423..4878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4510..4529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4720..4741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4995..5352
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4995..5186
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5018..5038)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5203..5338
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5403..6197
                     /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(5457..5476)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6067..6086
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6261..6309
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6350..6369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6469..6491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6529..6547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6615..6719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6720..6788,6789..7580)
                     /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(6938..6957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7751..8339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8240..8259
                     /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 cgacatggat gctgattctg cctccagttc tgaatcagag agaaacatca
     1321 ctatcatggc ttcagggaac acaggtggtg aaaaagatgg ccttcggaat agcactggac
     1381 ttggttccca aaacaagttt gtagttggta gcagcagcaa taatgtgggc catggaagta
     1441 gtactgggcc atggggtttt tcccatggag ccataataag cacatgtcag gtctctgtgg
     1501 atgctcctga aagcaaatct gaaagtagca acaatagaat gaatgcttgg ggcactgtaa
     1561 gttcttcatc aaatggaggg ttaaatccaa gcactttgaa ttcagctagc aaccatggtg
     1621 cctggccagt attagagaac aatggacttg ccctaaaagg gcctgtaggg agtggtagtt
     1681 ctggcattaa tattcagtgc agtactatag gccagatgcc taacaatcag agtattaact
     1741 ctaaagtgag tggtggttct acccatggta cctggggaag ccttcaggaa acttgtgaat
     1801 ctgaagtaag tggtacacag aaggtttcat tcagtggtca acctcaaaat attaccactg
     1861 aaatgactgg accaaataac actactaact ttatgacctc tagtttacca aactccggtt
     1921 cagtgcagaa taatgagctg cctagtagta acacaggggc ctggcgtgtg agcacaatga
     1981 atcatcctca gatgcaggct ccatcaggta tgaatggcac ttccctttct caccttagca
     2041 atggagagtc aaaaagtgga ggctcttatg gtactacatg gggtgcctat ggttctaatt
     2101 actctggaga caaatgttca ggccctaatg gccaagctaa tggtgacact gtgaatgcaa
     2161 ctctaatgca gcctggcgta aatggtccta tgggcactaa ctttcaagtt aacacaaaca
     2221 aaggaggtgg tgtgtgggaa tctggtgcag caaactccca gagtacatca tggggaagtg
     2281 gaaatggcgc aaattctgga ggaagtcgaa gaggatgggg aacccctgca caaaacactg
     2341 gcactaattt acccagcgtt gagtggaaca aactgcctag caatcagcat tccaatgata
     2401 gtgcaaatgg caatggtaag acgtttacaa atggatggaa atctactgag gaagaggatc
     2461 agggttctgc cacatctcag acaaatgagc aaagcagtgt gtgggccaaa acaggaggta
     2521 cagtggagag cgatggtagt acagaaagca ctggacgcct tgaggaaaaa ggaactgggg
     2581 aaagtcagag tagagacaga agaaaaattg atcagcacac attactccaa agcattgtaa
     2641 acagaactga cttagatcca cgtgtcctgt ccaactctgg ttggggacag actcctatta
     2701 agcagaatac tgcctgggat acagaaacat cacctagagg ggaacgaaag actgacaatg
     2761 ggacagaggc ctggggaagc tctgcaacac agacttttaa ctcaggggca tgtatagata
     2821 agactagccc taatggtaat gatacctcat ctgtatcagg gtggggcgat cccaaacctg
     2881 ctctgaggtg gggagattcc aaaggctcaa actgccaggg ggggtgggaa gatgattctg
     2941 ctgctacagg aatggtcaag agcaatcagt gggggaattg caaagaggag aaggctgcat
     3001 ggaatgactc gcaaaagaat aaacagggat ggggtgatgg acaaaaatca agccaagggt
     3061 ggtctgtttc tgccagtgat aactggggag aaacttcaag gaataaccat tggggtgagg
     3121 ccaataagaa atccagctca ggaggtagtg acagtgacag gtccgtttcc ggttggaacg
     3181 aacttggtaa aactagttct ttcacttggg gaaacaacat aaatccaaat aattcatcag
     3241 gatgggatga atcttctaaa cctactcctt cccagggatg gggagaccct ccaaagtcta
     3301 atcagtctct aggttgggga gattcgtcaa agccagtcag ctctccagac tggaacaagc
     3361 aacaagacat tgttggatct tggggaatcc caccagctac aggcaaacct cctggtacag
     3421 gctggctggg gggacctata ccagccccag caaaagaaga agaacccaca ggctgggagg
     3481 aaccatcccc agaatctata cgtcgcaaaa tggagattga tgatggaact tcagcttggg
     3541 gagatccaag caaatacaac tacaaaaatg tgaacatgtg gaacaaaaac gtcccaaatg
     3601 gcaacagccg ttcagaccag caagcacagg tacatcagct gctaacgcct gcaagtgcca
     3661 tctcaaacaa agaggcaagc agtggctctg gctggggtga gccctggggg gagccttcta
     3721 ctccagccac aactgtggat aatggtactt cagcatgggg taagcccata gacagtggtc
     3781 ccagctgggg ggaacccatt gctgcggcat ccagcacatc cacgtggggc tccagctctg
     3841 ttggtccaca agcattaagc aaatctgggc caaaatctat gcaagatggc tggtgtggtg
     3901 atgatatgcc attgcctgga aatcgcccca ctggctggga agaggaagag gatgtggaga
     3961 ttggaatgtg gaatagtaat tcatctcaag agcttaactc atctttaaat tggccaccat
     4021 atacaaagaa aatgtcatcg aagggtctga gtggcaaaaa aaggagaagg cggccgcttc
     4081 cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg agtttggaca
     4141 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4201 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4261 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     4321 atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga ggcccgcacc
     4381 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     4441 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4501 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4561 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4621 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4681 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4741 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4801 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4861 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4921 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     4981 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     5041 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5101 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5161 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5221 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5281 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5341 ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca
     5401 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5461 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5521 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5581 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5641 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5701 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5761 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5821 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5881 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5941 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6001 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6061 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6121 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6181 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6241 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     6301 ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt acaatctgct
     6361 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6421 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6481 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     6541 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6601 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6661 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6721 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6781 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6841 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6901 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6961 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     7021 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7081 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7141 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7201 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7261 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7321 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7381 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7441 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7501 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7561 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7621 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7681 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7741 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7861 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     8041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8341 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
     8401 ctcgacagat ct
//
