LOCUS       sequence-289041-        6721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6C_1248-1690-delMID2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6721)
  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 6721)
  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-delMID2_I
FEATURES             Location/Qualifiers
     source          1..6721
                     /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    2425..2546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2436..2455)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2490..2509
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2732..3187
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2819..2838)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3029..3050
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3304..3661
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3304..3495
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3327..3347)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3512..3647
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3712..4506
                     /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(3766..3785)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4376..4395
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4570..4618
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4659..4678)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4778..4800
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4838..4856)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4924..5028
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5029..5097,5098..5889)
                     /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(5247..5266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6060..6648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6549..6568
                     /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 ctggcagtag aaccagcagc tggctcgttc ttcgaaacct cactccccag attgatggtt
     1801 ctacactgcg gacattgtgt ttgcaacatg ggcctcttat cacattccac ctgaatctga
     1861 ctcaaggcaa tgctgtggtc cggtacagct ccaaggagga ggctgccaag gcccagaagt
     1921 ctctgcacat gtgcgtcctg ggaaacacta ccatcctggc cgagttcgct ggtgaagaag
     1981 aagtgaatcg cttcttagcc caaggccagg cgctgccacc cacttccagc tggcagtcca
     2041 gcagcgcgtc cagccagccg cggctcagcg cagcgggcag ctcccatggc ctggtacgca
     2101 gcgacgctgg ccactggaac gccccgtgcc tgggtggcaa ggggagcagt gagctgctgt
     2161 ggggcggggt gccccagtac tccagcagcc tgtggggccc gcccagcgcc gacgacagca
     2221 gggtgatagg cagccccacg ccgctaacca ccctgctgcc tggggacctg ctcagcgggg
     2281 agtccctgta ggcgctgcca tcatcagcac caggagagcc gacccctccc gggacccctc
     2341 ccggctgggc ggccccacag acccgctgga acccagcagc ggccgcttcc ctttagtgag
     2401 ggttaatgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     2461 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2521 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2581 ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     2641 tccgataagg atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     2701 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg
     2761 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2821 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2881 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2941 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3001 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3061 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3121 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3181 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3241 gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     3301 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3361 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3421 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3481 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3541 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3601 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3661 aaagcttgat tcttctgaca caacagtctc gaacttaagg ctagagccac catgattgaa
     3721 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3781 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3841 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3901 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3961 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4021 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4081 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4141 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4201 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4261 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4321 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4381 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4441 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4501 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4561 gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     4621 tcgatagcga taaggatccg cgtatggtgc actctcagta caatctgctc tgatgccgca
     4681 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4741 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4801 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4861 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4921 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4981 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5041 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5101 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5161 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5221 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5281 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5341 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5401 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5461 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5521 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5581 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5641 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5701 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5761 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5821 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5881 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5941 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6001 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6061 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6121 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6181 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6241 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6301 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6361 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6421 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6481 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6541 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6601 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6661 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatggc tcgacagatc
     6721 t
//
