LOCUS       sequence-288739-        7934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6A-Cterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7934)
  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 7934)
  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-C-term_F
FEATURES             Location/Qualifiers
     source          1..7934
                     /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             1191..1217
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2660..2671
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     polyA_signal    3638..3759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3649..3668)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3703..3722
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3945..4400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4032..4051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4242..4263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4517..4874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4517..4708
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4540..4560)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4725..4860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4925..5719
                     /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(4979..4998)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5589..5608
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5783..5831
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5872..5891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5991..6013
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6051..6069)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6137..6241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6242..6310,6311..7102)
                     /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(6460..6479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7273..7861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7762..7781
                     /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 agaattcacg cgtgatggct tacccatacg
     1201 atgttccaga ttacgctctg agtggcaaaa aaaggagaag ggaaagggga atgatgaaag
     1261 gtggaaacaa acaagaagaa gcgtggataa atccatttgt taaacagttt tcaaacatca
     1321 gtttttcgag agactcacca gaggaaaatg tacaaagcaa taagatggac ctttctggag
     1381 gaatgttaca agacaaacga atggagatag ataaacatag cctaaatatt ggtgattaca
     1441 atcgaacggt cgggaaaggc cctggttctc ggcctcagat ttccaaagag tcttccatgg
     1501 agcgcaatcc ttattttgat aaggatggca ttgtagcaga tgaatcccaa aacatgcagt
     1561 ttatgtccag tcaaagcatg aagcttcccc cttcaaatag tgcactacct aaccaggccc
     1621 ttggctccat agcagggctg ggtatgcaaa acttgaattc tgttagacag aatggcaatc
     1681 ccagtatgtt tggtgttgga aacacagcag cacaaccccg gggcatgcag cagcctccag
     1741 cacaacctct tagttcatct cagcctaatc tccgtgctca agtgcctcct ccattactct
     1801 cccctcaggt tccagtttca ttgctgaagt atgcaccaaa caacggtggc ctgaatccac
     1861 tctttggccc tcaacaggta gccatgctga accagctatc ccagctaaac cagctttctc
     1921 agatctccca gttacagcga ttgttagcgc agcagcaaag ggcgcagagt cagagaagcg
     1981 tgccttctgg gaaccggccg cagcaagacc agcagggtcg acctcttagt gtgcagcagc
     2041 aaatgatgca acaatctcgt caacttgatc caaacctgtt ggtgaagcag cagactccac
     2101 catctcagca gcagccactc catcagccag ccatgaagtc tttccttgac aatgtcatgc
     2161 cccacactac acctgagctg caaaaagggc catcaccaat aaatgctttc agcaacttcc
     2221 ctataggctt gaactcaaac ttgaatgtaa atatggatat gaacagtatt aaagagccac
     2281 agtcaagact aaggaagtgg acgacagtgg acagcatttc tgtgaacaca tctttggatc
     2341 aaaactccag caaacatggt gctatttcaa gtggtttcag gctggaagag tctccatttg
     2401 ttccctatga ctttatgaac agcagtactt caccagccag tcctccaggt tcaataggag
     2461 atggctggcc acgtgccaaa tcgcctaacg gctctagcag tgttaattgg ccaccagaat
     2521 ttcgtcctgg tgagccatgg aaaggttatc caaacattga ccctgaaact gacccttacg
     2581 tcactcctgg cagtgtcata aacaatcttt caattaatac tgtgcgggaa gttgaccacc
     2641 tcagggacag gaacagtggg tcatcctcat ccttgaacac cacgctgcct tcaactagtg
     2701 cctggtcatc cattcgtgcc tccaactaca acgttcccct cagcagtaca gcacaaagca
     2761 cttcagccag aaatagtgat tccaaattga catggtctcc tggttcagtt acaaacacct
     2821 ctctggctca tgagctgtgg aaggtccctt tgccacctaa aaacatcact gctccgtccc
     2881 gcccacctcc gggactgact ggtcagaagc cacccttgtc tacgtgggat aattctcccc
     2941 ttcgtatagg tggaggatgg ggaaattctg acgccagata taccccaggt tccagctggg
     3001 gtgagagcag ctcagggaga ataacaaatt ggcttgttct aaaaaacctt acacctcaga
     3061 tcgatggctc aactctgcgc actctgtgca tgcagcacgg cccgctgatc acattccacc
     3121 tgaacctccc tcacggaaat gctctggtcc gctacagttc aaaagaagag gtagtggagg
     3181 cacaaaagtc tctgcacatg tgtgtactgg ggaacactac tattcttgct gagtttgcca
     3241 gtgaagagga gatcagtcgt ttctttgcac aaagccagtc tctgacccct tctcccggct
     3301 ggcagtctct cgggtccagc cagagccggc tgggctccct cgactgttcc cactcattct
     3361 ccagccggac cgatctcaat cactggaatg gtgctgggct gtcgggaact aactgtggag
     3421 accttcacgg cacttcactc tgggggaccc cgcattattc cacaagcctg tggggtcccc
     3481 caagcagcag cgacccccga ggaattagca gcccatctcc cattaacgct tttctttctg
     3541 ttgaccacct gggtgggggt ggagagtcca tgtaatctag agtcgacccg ggcggccgct
     3601 tccctttagt gagggttaat gcttcgagca gacatgataa gatacattga tgagtttgga
     3661 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3721 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3781 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     3841 aaatgtggta aaatccgata aggatcgatc cgggctggcg taatagcgaa gaggcccgca
     3901 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     3961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4381 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4441 tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa
     4501 cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4561 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4621 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4681 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4741 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4801 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4861 taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta aggctagagc
     4921 caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4981 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5041 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5101 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5161 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5221 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5281 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5341 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5401 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5461 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5521 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5581 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5641 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5701 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5761 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     5821 ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg tgcactctca gtacaatctg
     5881 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5941 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     6001 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     6061 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     6121 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6181 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6241 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6301 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6361 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6421 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6481 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6541 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6601 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6661 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6721 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6781 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6841 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6901 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6961 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7021 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7081 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7141 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     7201 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7261 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7321 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7381 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7441 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7501 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7561 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7621 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7681 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7741 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7801 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7861 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7921 ggctcgacag atct
//
