LOCUS       sequence-289042-        6742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6C_1248-1690-delRRM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6742)
  AUTHORS   Eulalio A, Helms S, Fritzsch C, Fauser M, Izaurralde E
  TITLE     A C-terminal silencing domain in GW182 is essential for miRNA 
            function.
  JOURNAL   RNA. 2009 Jun;15(6):1067-77. doi: 10.1261/rna.1605509. Epub 2009 Apr
            21.
  PUBMED    19383769
REFERENCE   2  (bases 1 to 6742)
  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: 
            "10.1261/rna.1605509"; journalName: "RNA"; date: "2009-06"; volume: 
            "15"; issue: "6"; pages: "1067-77"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsTNRC6C_1248-1690-delRRM_I
FEATURES             Location/Qualifiers
     source          1..6742
                     /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    2446..2567
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2457..2476)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2511..2530
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2753..3208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2840..2859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3050..3071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3325..3682
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3325..3516
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3348..3368)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3533..3668
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3733..4527
                     /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(3787..3806)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4397..4416
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4591..4639
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4680..4699)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4799..4821
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4859..4877)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4945..5049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5050..5118,5119..5910)
                     /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(5268..5287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6081..6669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6570..6589
                     /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 gaattccatc
     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 acctcaggac
     2041 ccacttccag ctggcagtcc agcagcgcgt ccagccagcc gcggctcagc gcagcgggca
     2101 gctcccatgg cctggtacgc agcgacgctg gccactggaa cgccccgtgc ctgggtggca
     2161 aggggagcag tgagctgctg tggggcgggg tgccccagta ctccagcagc ctgtggggcc
     2221 cgcccagcgc cgacgacagc agggtgatag gcagccccac gccgctaacc accctgctgc
     2281 ctggggacct gctcagcggg gagtccctgt aggcgctgcc atcatcagca ccaggagagc
     2341 cgacccctcc cgggacccct cccggctggg cggccccaca gacccgctgg aacccagcag
     2401 cggccgcttc cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg
     2461 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2521 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2581 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     2641 acctctacaa atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga
     2701 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     2761 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2821 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2881 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2941 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3001 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3061 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3121 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3181 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     3241 gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga
     3301 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     3361 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3421 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3481 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3541 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3601 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3661 tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag
     3721 gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3781 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3841 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3901 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3961 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4021 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4081 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4141 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4201 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4261 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4321 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4381 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4441 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4501 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4561 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     4621 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt
     4681 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4741 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4801 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4861 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4921 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4981 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5041 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5101 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5161 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5221 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5281 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5341 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5401 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5461 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5521 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5581 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5641 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5701 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5761 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5821 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5881 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5941 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6001 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6061 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6121 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6181 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6241 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6301 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6361 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6421 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6481 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6541 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6601 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6661 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6721 gctcacatgg ctcgacagat ct
//
