LOCUS       sequence-288746-        7831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6Biso1-Cterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7831)
  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 7831)
  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-C-term_F
FEATURES             Location/Qualifiers
     source          1..7831
                     /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    3535..3656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3546..3565)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3600..3619
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3842..4297
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3929..3948)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4139..4160
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4414..4771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4414..4605
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4437..4457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4622..4757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4822..5616
                     /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(4876..4895)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5486..5505
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5680..5728
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5769..5788)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5888..5910
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5948..5966)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6034..6138
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6139..6207,6208..6999)
                     /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(6357..6376)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7170..7758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7659..7678
                     /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 cgaccaagga ggaaagaaac aaatgaagtg ctcactcaaa ggaggaaaca
     1321 atgattcatg gatgaatcct cttgccaaac agttttcaaa tatgggattg ctgagtcaga
     1381 ctgaagataa tccaagcagc aaaatggatt tgtctgtagg aagcctttca gataaaaaat
     1441 ttgatgtgga caagcgagcg atgaatctcg gggattttaa tgatatcatg aggaaggatc
     1501 gatctgggtt ccgtccacct aattccaaag acatgggaac cacagatagt gggccttatt
     1561 ttgagaaggg cggtagtcat ggtttgtttg gaaacagcac agcacaatcg agaggtctgc
     1621 acacacccgt gcagccacta aattcttctc ccagtctccg ggcgcaagtg cctccccagt
     1681 ttatttcccc ccaggtttct gcctcaatgc tcaagcagtt tcccaacagt ggcctgagtc
     1741 caggtctttt caatgtgggg ccccagttat ctcctcaaca aattgccatg ctgagccagc
     1801 ttccacaaat tccccagttt cagttggcat gtcagcttct cttgcagcag cagcaacagc
     1861 agcagttgtt acagaaccag agaaagattt ctcaagctgt acgccaacag caagagcagc
     1921 agctggctcg aatggtgagt gcactgcagc agcagcagca gcagcagcag aggcagccag
     1981 gcatgaagca ctcgccctct catcctgttg ggcccaagcc gcatctggac aacatggtac
     2041 ccaacgcatt gaatgtgggg ctcccagacc ttcaaaccaa agggccaata cctggatatg
     2101 gttctggctt cagctctggc ggcatggact atggcatggt tggtgggaag gaggctggaa
     2161 ccgagtctcg ctttaaacag tggacctcca tgatggaggg gctgccctct gtagccacac
     2221 aggaagccaa tatgcacaaa aatggcgcta tagtggcccc tggtaaaacc cggggagggt
     2281 caccgtacaa ccagtttgat atcatccctg gtgacacact gggtggccat acgggtcctg
     2341 ctggtgatag ctggttacct gccaaatctc caccaacaaa taaaatcgga agtaaatcca
     2401 gcaatgccag ttggcctcca gaattccaac caggagtgcc atggaaaggt atccaaaaca
     2461 ttgaccctga atctgacccc tatgtcaccc caggaagtgt gctggggggt acagccacat
     2521 ctcccattgt agatactgac caccaactgc tgcgggataa caccacaggg tctaattctt
     2581 ccctcaacac ctcgctgcct tcacctggtg cctggcccta cagtgcctct gacaactcct
     2641 ttaccaacgt tcatagcact tcagcaaagt tccctgatta caaatcaaca tggtccccag
     2701 atcccatagg acacaacccc actcatctct ccaacaagat gtggaaaaac catatttcct
     2761 ccaggaacac tacaccgctg ccccgcccac ctcctggtct gaccaacccc aaaccatcat
     2821 ctccctggag cagcacagca ccccgatcag tcagggggtg ggggacacag gactcacggc
     2881 tcgcctcggc ctctacctgg agtgatggtg gctcagttcg tcctagttac tggctggttc
     2941 ttcacaatct caccccacag attgatgggt caaccttgag aacgatctgc atgcagcatg
     3001 gcccactgct gacattccat ctgaatctaa cccagggcac tgccctgatc cgatacagca
     3061 ccaaacagga ggcggccaag gcccaaactg cactgcacat gtgtgtgttg ggaaacacta
     3121 ccatccttgc tgagtttgcc actgatgatg aagtcagccg ctttctggca caagctcagc
     3181 cccctacacc tgcagcaacc ccaagtgcgc cagctgcggg gtggcagtcg ctggagaccg
     3241 gccagaacca gtcagatccc gtgggacctg ctctgaatct ttttggtggg tccactgggc
     3301 tcgggcagtg gagcagcagc gctggtggca gcagcggggc cgatcttgct ggcgcttcat
     3361 tgtgggggcc cccaaactat tcttctagct tatggggagt cccaacggtg gaagatcccc
     3421 ataggatggg cagccctgct cctttactac ctggtgacct tctgggagga gggtcggatt
     3481 caatctgagc ggccgcttcc ctttagtgag ggttaatgct tcgagcagac atgataagat
     3541 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3601 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3661 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     3721 gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa
     3781 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3841 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3901 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3961 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4021 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4081 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4141 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4201 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4261 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     4321 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     4381 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     4441 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4501 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4561 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4621 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4681 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4741 aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc
     4801 gaacttaagg ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc
     4861 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4921 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4981 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5041 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5101 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5161 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5221 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5281 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5341 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5401 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5461 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5521 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5581 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5641 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     5701 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc
     5761 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     5821 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5881 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5941 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6001 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6061 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6121 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6181 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6241 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6301 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6361 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6421 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6481 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6541 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6601 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6661 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6721 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6781 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6841 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6901 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6961 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7021 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7081 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7141 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7201 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7261 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7321 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7381 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7441 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7501 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7561 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7621 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7681 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7741 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7801 tggccttttg ctcacatggc tcgacagatc t
//
