LOCUS       sequence-357212-        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6A_1242-1709-F1359A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  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 6955)
  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-HsTNRC6A_1242-1709-F1359A_K
FEATURES             Location/Qualifiers
     source          1..6955
                     /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             1179..1205
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1697..1708
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     polyA_signal    2659..2780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2670..2689)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2724..2743
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2966..3421
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3053..3072)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3263..3284
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3538..3895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3538..3729
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3561..3581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3746..3881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3946..4740
                     /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(4000..4019)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4610..4629
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4804..4852
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4893..4912)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5012..5034
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5072..5090)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5158..5262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5263..5331,5332..6123)
                     /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(5481..5500)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6294..6882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6783..6802
                     /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 agatggctta cccatacgat gttccagatt
     1201 acgctacacc tgagctgcaa aaagggccat caccaataaa tgctttcagc aacttcccta
     1261 taggcttgaa ctcaaacttg aatgtaaata tggatatgaa cagtattaaa gagccacagt
     1321 caagactaag gaagtggacg acagtgggca gcatttctgt gaacacatct ttggatcaaa
     1381 actccagcaa acatggtgct atttcaagtg gtttcaggct ggaagagtct ccatttgttc
     1441 cctatgactt tatgaacagc agtacttcac cagccagtcc tccaggttca ataggagatg
     1501 gctggccacg tgccaaatcg cctaacggct ctagcagtgt taattggcca ccagaagctc
     1561 gtcctggtga gccatggaaa ggttatccaa acattgaccc tgaaactgac ccttacgtca
     1621 ctcctggcag tgtcataaac aatctttcaa ttaatactgt gcgggaagtt gaccacctca
     1681 gggacaggaa cagtgggtca tcctcatcct tgaacaccac gctgccttca actagtgcct
     1741 ggtcatccat tcgtgcctcc aactacaacg ttcccctcag cagtacagca caaagcactt
     1801 cagccagaaa tagtgattcc aaattgacat ggtctcctgg ttcagttaca aacacctctc
     1861 tggctcatga gctgtggaag gtccctttgc cacctaaaaa catcactgct ccgtcccgcc
     1921 cacctccggg actgactggt cagaagccac ccttgtctac gtgggataat tctccccttc
     1981 gtataggtgg aggatgggga aattctgacg ccagatatac cccagctgga acctggggtg
     2041 agagcagctc agggagaata acaaattggc ttgttctaaa aaaccttaca cctcagatcg
     2101 atggctcaac tctgcgcact ctgtgcatgc agcacggccc gctgatcaca ttccacctga
     2161 acctccctca cggaaatgct ctggtccgct acagttcaaa agaagaggta gtggaggcac
     2221 aaaagtctct gcacatgtgt gtactgggga acactactat tcttgctgag tttgccagtg
     2281 aagaggagat cagtcgtttc tttgcacaaa gccagtctct gaccccttct cccggctggc
     2341 agtctctcgg gtccagccag agccggctgg gctccctcga ctgttcccac tcattctcca
     2401 gccggaccga tctcaatcac tggaatggtg ctgggctgtc gggaactaac tgtggagacc
     2461 ttcacggcac ttcactctgg gggaccccgc attattccac aagcctgtgg ggtcccccaa
     2521 gcagcagcga cccccgagga attagcagcc catctcccat taacgctttt ctttctgttg
     2581 accacctggg tgggggtgga gagtccatgt aagcggccgc ttccctttag tgagggttaa
     2641 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2701 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2761 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2821 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     2881 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2941 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3001 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3061 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3121 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3181 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3241 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3301 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3361 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3421 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     3481 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     3541 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3601 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3661 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3721 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3781 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3841 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3901 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     3961 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4021 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4081 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4141 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4201 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4261 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4321 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4381 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4441 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4501 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4561 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4621 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4681 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4741 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     4801 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     4861 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4921 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4981 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5041 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5101 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5161 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5221 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5281 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5341 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5401 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5461 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5521 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5581 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5641 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5701 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5761 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5821 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5881 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5941 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6001 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6061 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6121 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6181 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6241 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6301 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6361 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6421 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6481 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6541 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6601 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6661 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6721 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6781 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6841 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6901 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatct
//
