LOCUS       sequence-355556-        8395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6C-Nterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  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 8395)
  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-HsTNRC6C-N-term_F
FEATURES             Location/Qualifiers
     source          1..8395
                     /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"
     CDS             3554..3571
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4099..4220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4110..4129)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4164..4183
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4406..4861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4493..4512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4703..4724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4978..5335
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4978..5169
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5001..5021)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5186..5321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5386..6180
                     /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(5440..5459)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6050..6069
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6244..6292
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6333..6352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6452..6474
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6512..6530)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6598..6702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6703..6771,6772..7563)
                     /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(6921..6940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7734..8322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8223..8242
                     /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 cgacatggct acagggagtg cccagggcaa cttcactgga cataccaaga
     1321 agacaaacgg caataatggc accaatggcg cactcgtcca aagcccttct aatcagagtg
     1381 cccttggagc agggggagcg aacagtaatg gaagtgcggc cagagtgtgg ggtgtagcca
     1441 caggctccag ctctggcctg gctcactgct ctgtcagtgg tggggatgga aaaatggaca
     1501 ctatgattgg agatgggaga agtcagaatt gctggggtgc ttccaactcc aatgctggca
     1561 ttaatcttaa ccttaatcct aatgccaacc cagctgcctg gcctgtactt ggacatgaag
     1621 gaaccgtggc gacaggcaac ccttccagta tttgcagtcc agtcagtgcc ataggtcaaa
     1681 atatgggcaa ccagaacggg aacccaacag gcactttagg tgcttgggga aacttgctgc
     1741 cacaagagag cacagaacca caaacgtcca cttctcagaa tgtgtctttc agcgcacaac
     1801 ctcagaacct taacactgat ggaccaaata acactaaccc catgaactct tcacccaacc
     1861 ctatcaatgc aatgcagaca aatggactgc caaactgggg catggctgtt ggtatggggg
     1921 ccatcatccc gccccacctg caaggccttc ctggtgctaa tggatcatca gtttctcaag
     1981 tcagtggggg cagtgctgaa ggaataagca attctgtgtg gggactgtcc ccaggtaacc
     2041 ctgccacagg aaatagcaat tctgggttca gtcaggggaa tggagacact gtgaactcag
     2101 cattaagtgc taaacaaaat ggatccagca gtgctgtgca aaaggaagga agtggaggaa
     2161 atgcttggga ttcaggacct cctgctggtc ctggaatact cgcctgggga aggggcagtg
     2221 gcaacaatgg cgttggtaat atccattcag gagcttgggg ccaccccagc cgaagcacct
     2281 ctaacggtgt gaatggggaa tggggaaagc ccccaaacca gcattccaac agtgacatca
     2341 atgggaaagg atcaacaggg tgggagagtc ctagtgtcac cagccagaac cctaccgtac
     2401 agcctggtgg tgaacacatg aactcctggg ccaaagcggc atcttctgga actacagcaa
     2461 gtgaaggaag tagtgatggt tctggcaacc acaatgaagg aagcactggg agggaaggaa
     2521 cgggagaagg ccgaaggcga gataaaggga ttatagacca agggcacatc cagttgccaa
     2581 ggaatgatct tgacccaaga gttctgtcta atactggttg gggacagact cctgtaaagc
     2641 aaaacactgc ctgggaattt gaagaatccc ctaggtctga aaggaaaaat gacaatggga
     2701 cagaggcctg gggttgtgca gctactcagg cttcaagctc aggggggaag aacgatgggt
     2761 ccatcatgaa cagtacaaat acctcttcag tatctgggtg ggtcaacgcg ccacctgccg
     2821 ctgtgccagc aaacacaggt tggggagaca gcaacaacaa agcgccaagt ggcccggggg
     2881 tttgggggga ctcgataagc tctactgctg ttagtactgc tgctgctgcc aagagtggcc
     2941 atgcttggag tggggccgca aatcaggagg acaagtcacc cacctggggt gagcctccaa
     3001 agcccaaatc ccaacactgg ggagatggac aaagatcaaa tccagcctgg agtgcaggag
     3061 ggggagattg ggcagattca tcgtctgtcc ttggacactt gggggatggg aaaaaaaatg
     3121 gatctggatg ggatgctgac agtaataggt cagggtctgg ttggaatgac accacgagat
     3181 ctgggaacag tggctggggc aacagcacaa atacaaaggc caatccaggt acaaactggg
     3241 gggagacttt aaaacctggc ccccaacaga actgggctag caaaccccaa gacaacaatg
     3301 tgagtaactg gggaggagct gcttctgtga aacagacagg aacagggtgg atcggggggc
     3361 cggtaccggt caaacagaag gacagcagtg aagcaactgg ctgggaagaa ccctctccac
     3421 cgtccattcg ccgcaaaatg gaaattgatg atggtacctc agcttggggg gacccaagca
     3481 actataacaa taaaactgta aacatgtggg atagaaacaa cccggtcatc cagagcagta
     3541 ccacgaccaa taccaccacc accaccacca ctaccacgag caacaccaca cacagggtcg
     3601 agacgccgcc cccgcaccag gctggtactc agctgaatcg atcaccgttg cttggtccag
     3661 gtaggaaagt ttcatcaggc tggggagaaa tgcctaatgt tcactcaaag actgaaaact
     3721 cttggggaga accatcctcc ccttctaccc tggtggataa tggcacagca gcatggggga
     3781 agccacccag cagtggcagc gggtggggag atcaccctgc cgagccgccg gtggcatttg
     3841 gaagagctgg cgcacctgtt gctgcctcag ccctgtgcaa accagcttca aaatctatgc
     3901 aagaaggctg gggcagtggt ggggatgaaa tgaacctcag taccagccag tgggaggatg
     3961 aagaagggga cgtgtggaat aatgctgctt cccaagaaag cacctcctcc tgcagctcct
     4021 gggggaacgc ccccaaaaaa ggacttcaaa aggcggccgc ttccctttag tgagggttaa
     4081 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4141 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4201 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4261 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     4321 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4381 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4441 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4501 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4561 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4621 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4681 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4741 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4801 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     4861 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     4921 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     4981 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5041 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5101 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5161 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5221 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5281 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5341 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     5401 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5461 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5521 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5581 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5641 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5701 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5761 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5821 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5881 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5941 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6001 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6061 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6121 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6181 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     6241 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     6301 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6361 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6421 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6481 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6541 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6601 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6661 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6721 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6781 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6841 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6901 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6961 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7021 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7081 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7141 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7201 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7261 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7321 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7381 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7441 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7501 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7561 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7621 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7681 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7741 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7801 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7861 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7921 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7981 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8041 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8101 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8161 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8221 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8281 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8341 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatct
//
