LOCUS       sequence-289192-        7012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6C_1248-1690-F1389A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7012)
  AUTHORS   Huntzinger E, Braun JE, Heimstadt S, Zekri L, Izaurralde E
  TITLE     Two PABPC1-binding sites in GW182 proteins promote miRNA-mediated 
            gene silencing.
  JOURNAL   EMBO J. 2010 Dec 15;29(24):4146-60. doi: 10.1038/emboj.2010.274
  PUBMED    21063388
REFERENCE   2  (bases 1 to 7012)
  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"; doi: 
            "10.1038/emboj.2010"; journalName: "EMBO J"; date: "2010-12-15- 15";
            volume: "29"; issue: "24"; pages: "4146-60"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsTNRC6C_1248-1690-F1389A_K
FEATURES             Location/Qualifiers
     source          1..7012
                     /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    2716..2837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2727..2746)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2781..2800
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3023..3478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3110..3129)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3320..3341
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3595..3952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3595..3786
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3618..3638)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3803..3938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4003..4797
                     /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(4057..4076)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4667..4686
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4861..4909
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4950..4969)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5069..5091
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5129..5147)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5215..5319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5320..5388,5389..6180)
                     /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(5538..5557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6351..6939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6840..6859
                     /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 cctgacctgc
     1261 agaccaaaga gcagcagtct tcacccaaca cctttgctcc ttaccctctc gctggactga
     1321 acccaaacat gaatgtcaac agcatggaca tgaccggtgg cttgtcggtg aaggacccat
     1381 cccagtccca gtcacgcctc ccccagtgga cgcaccccaa ctccatggat aacttgccca
     1441 gtgccgcttc ccccctggag cagaacccta gcaagcatgg tgctatccct ggaggtctaa
     1501 gcattgggcc tccaggtaag tcctccattg atgactccta tggccggtac gatttaatcc
     1561 agaacagtga gtcaccagcc agtcctcccg tagctgttcc ccatagctgg tcacgtgcca
     1621 aatctgacag tgataaaatc tcaaatggct ctagcatcaa ctggccccca gaagcccatc
     1681 cgggagttcc atggaaagga ctgcagaata ttgaccctga gaatgaccct gacgtcactc
     1741 ctggcagtgt ccccactggg cctaccatca acaccaccat ccaggatgtc aaccgctacc
     1801 tcctcaagag tggaggtaaa ctgtcagaca tcaaatcgac gtggtcctct ggccctacct
     1861 cccacacgca agcctctctg tctcatgaac tatggaaggt gcccagaaac agtactgcac
     1921 ccacgaggcc acctccaggg ttaaccaatc ccaagccctc ctccacctgg ggtgccagcc
     1981 ccctcggctg gaccagctcc tactcctcgg gttctgcctg gagcaccgac acctcaggaa
     2041 gaaccagcag ctggctcgtt cttcgaaacc tcactcccca gattgatggt tctacactgc
     2101 ggacattgtg tttgcaacat gggcctctta tcacattcca cctgaatctg actcaaggca
     2161 atgctgtggt ccggtacagc tccaaggagg aggctgccaa ggcccagaag tctctgcaca
     2221 tgtgcgtcct gggaaacact accatcctgg ccgagttcgc tggtgaagaa gaagtgaatc
     2281 gcttcttagc ccaaggccag gcgctgccac ccacttccag ctggcagtcc agcagcgcgt
     2341 ccagccagcc gcggctcagc gcagcgggca gctcccatgg cctggtacgc agcgacgctg
     2401 gccactggaa cgccccgtgc ctgggtggca aggggagcag tgagctgctg tggggcgggg
     2461 tgccccagta ctccagcagc ctgtggggcc cgcccagcgc cgacgacagc agggtgatag
     2521 gcagccccac gccgctaacc accctgctgc ctggggacct gctcagcggg gagtccctgt
     2581 aggcgctgcc atcatcagca ccaggagagc cgacccctcc cgggacccct cccggctggg
     2641 cggccccaca gacccgctgg aacccagcag cggccgcttc cctttagtga gggttaatgc
     2701 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2761 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2821 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2881 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atccgataag
     2941 gatcgatccg ggctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3001 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3061 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3121 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3181 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3241 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3301 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3361 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3421 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     3481 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     3541 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     3601 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3661 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3721 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3781 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3841 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3901 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga
     3961 ttcttctgac acaacagtct cgaacttaag gctagagcca ccatgattga acaagatgga
     4021 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4081 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4141 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4201 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4261 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4321 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4381 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4441 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4501 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4561 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4621 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4681 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4741 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4801 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     4861 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     4921 ataaggatcc gcgtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4981 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5041 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5101 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5161 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5521 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6001 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6481 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6961 ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat ct
//
