LOCUS       sequence-194505-        6513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of HA-tagged human TNRC6C CED in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6513)
  AUTHORS   Chekulaeva M, Mathys H, Zipprich JT, Attig J, Colic M, Parker R, 
            Filipowicz W
  TITLE     miRNA repression involves GW182-mediated recruitment of CCR4-NOT 
            through conserved W-containing motifs.
  JOURNAL   Nat Struct Mol Biol. 2011 Oct 7;18(11):1218-26. doi: 
            10.1038/nsmb.2166.
  PUBMED    21984184
REFERENCE   2  (bases 1 to 6513)
  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/nsmb"; 
            journalName: "Nat Struct Mol Biol"; date: "2011-10-7- 7"; volume: 
            "18"; issue: "11"; pages: "1218-26"
COMMENT     Sequence Label: pMCh-1016
FEATURES             Location/Qualifiers
     source          1..6513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..291
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(408..863)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(545..566)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     757..776
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1049..1170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1086..1105)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1140..1159
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2236..2262)
                     /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"
     CDS             complement(2242..2268)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2302..2320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2363..2382)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2365..2497
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(2637..2661)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2658..2869)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2687..2707)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2870..3249
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3460..4048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3540..3559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4219..5010,5011..5079))
                     /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     4842..4861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5080..5184)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5252..5270
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5308..5330)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5430..5449
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    5490..5538
                     /note="synthetic polyadenylation signal"
     CDS             complement(5602..6396)
                     /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(5713..5732)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6323..6342
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6447..6513,1..291))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6461..6513,1..83)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct
      301 aaccaagttc ctctttcaga ggttatttca ggccatggtg ctgcgcagat ccgcgtatgc
      361 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt
      421 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
      481 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
      541 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
      601 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
      661 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
      721 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
      781 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
      841 taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg
      901 cgatcggtgc gggcctcttc gctattacgc cagcccggat cgatccttat cggattttac
      961 cacatttgta gaggttttac ttgctttaaa aaacctccca catctccccc tgaacctgaa
     1021 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1081 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1141 tggtttgtcc aaactcatca atgtatctta tcatgtctgc tcgaagcatt aaccctcact
     1201 aaagggaagc ggccgcggct ctcctggtgc tgatgatggc agcgcctaca gggactcccc
     1261 gctgagcagg tccccaggca gcagggtggt tagcggcgtg gggctgccta tcaccctgct
     1321 gtcgtcggcg ctgggcgggc cccacaggct gctggagtac tggggcaccc cgccccacag
     1381 cagctcactg ctccccttgc cacccaggca cggggcgttc cagtggccag cgtcgctgcg
     1441 taccaggcca tgggagctgc ccgctgcgct gagccgcggc tggctggacg cgctgctgga
     1501 ctgccagctg gaagtgggtg gcagcgcctg gccttgggct aagaagcgat tcacttcttc
     1561 ttcaccagcg aactcggcca ggatggtagt gtttcccagg acgcacatgt gcagagactt
     1621 ctgggccttg gcagcctcct ccttggagct gtaccggacc acagcattgc cttgagtcag
     1681 attcaggtgg aatgtgataa gaggcccatg ttgcaaacac aatgtccgca gtgtagaacc
     1741 atcaatctgg ggagtgaggt ttcgaagaac gagccagctg ctggttcttc ctgaggtgtc
     1801 ggtgctccag gcagaacccg aggagtagga gctggtccag ccgagggggc tggcacccca
     1861 ggtggaggag ggcttgggat tggttaaccc tggaggtggc ctcgtgggtg cagtactgtt
     1921 tctgggcacc ttccatagtt catgagacag agaggcttgc gtgtgggagg tagggccaga
     1981 ggaccacgtc gatttgatgt ctgacagttt acctccactc ttgaggaggt agcggttgac
     2041 atcctggatg gtggtgttga tggtaggccc agtggggaca ctgccaggag tgacgtcagg
     2101 gtcattctca gggtcaatat tctgcagtcc tttccatgga actcccggat ggaattctgg
     2161 gggccagttg atgctagagc catttgagat tttatcactg tcagatttgg cacggaattc
     2221 gacgggatcc gcgtcgagtg aagcgtaatc tggaacatca tatgggtagg ccatggtgta
     2281 agcttgggca gcctcgacta gcctatagtg agtcgtatta agtactctag ccttaagagc
     2341 tgtaattgaa ctgggagtgg acacctgtgg agagaaaggc aaagtggatg tcagtaagac
     2401 caataggtgc ctatcagaaa cgcaagagtc ttctctgtct cgacaagccc agtttctatt
     2461 ggtctcctta aacctgtctt gtaaccttga tacttacctg cccagtgcct cacgaccaac
     2521 ttctgcaagg ctaccttaag agagtcactg cagcttaagt tcgagactgt tgtgtcagaa
     2581 gcactgactg cgttagcaat ttaactgtga taaactaccg caataaagct tctagtgatc
     2641 tgacggttca ctaaacgagc tctgcttata tagacctccc accgtacacg cctaccgccc
     2701 atttgcgtca acggggcggg cgatcgcagt tgttacgaca ttttggaaag tcccgttgat
     2761 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     2821 agtcaaaccg ctatccacgc ccattggtgt actgccaaaa ccgcatcacc atggtaatag
     2881 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccgtaaggtc atgtactggg
     2941 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgga cttggcatat
     3001 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     3061 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     3121 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     3181 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     3241 atcaatgcca acatggcggt catattggac atgagccaat ataaatgtac atattatgat
     3301 atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata taaccaatga
     3361 ataatatggc taatggccaa tattgaagat ctgtcgagcc atgtgagcaa aaggccagca
     3421 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3481 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3541 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3601 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3661 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3721 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3781 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3841 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3901 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3961 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4021 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4081 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4141 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4201 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4261 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4321 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4381 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4441 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4501 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4561 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4621 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4681 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4741 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4801 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4861 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4921 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4981 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5041 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5101 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5161 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5221 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5281 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5341 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5401 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5461 ccatacgcgg atccttatcg ctatcgattc acacaaaaaa ccaacacaca gatgtaatga
     5521 aaataaagat attttattgc ggccatcgtg atggcaggtt gggcgtcgct tggtcggtca
     5581 tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     5641 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     5701 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     5761 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     5821 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct
     5881 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     5941 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     6001 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     6061 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     6121 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     6181 cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag
     6241 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     6301 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     6361 cggagaacct gcgtgcaatc catcttgttc aatcatggtg gctctagcct taagttcgag
     6421 actgttgtgt cagaagaatc aagctttttg caaaagccta ggcctccaaa aaagcctcct
     6481 cactacttct ggaatagctc agaggccgag gcg
//
