LOCUS       sequence-288954-        6951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6A_1242-1709.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6951)
  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 6951)
  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-HsTNRC6A_1242-1709_H
FEATURES             Location/Qualifiers
     source          1..6951
                     /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)"
     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      2962..3417
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3049..3068)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3259..3280
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3534..3891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3534..3725
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3557..3577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3742..3877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3942..4736
                     /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(3996..4015)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4606..4625
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4800..4848
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4889..4908)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5008..5030
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5068..5086)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5154..5258
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5259..5327,5328..6119)
                     /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(5477..5496)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6290..6878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6779..6798
                     /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 acagtggaca gcatttctgt gaacacatct ttggatcaaa
     1381 actccagcaa acatggtgct atttcaagtg gtttcaggct ggaagagtct ccatttgttc
     1441 cctatgactt tatgaacagc agtacttcac cagccagtcc tccaggttca ataggagatg
     1501 gctggccacg tgccaaatcg cctaacggct ctagcagtgt taattggcca ccagaatttc
     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 cccaggttcc agctggggtg
     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 taaacaagta acaacaattg cattcatttt atgtttcagg ttcaggggga
     2821 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg
     2881 atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2941 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3001 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3061 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3121 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3181 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3241 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3301 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3361 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     3421 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     3481 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     3541 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3661 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3721 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3781 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3841 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat
     3901 tcttctgaca caacagtctc gaacttaagg ctagagccac catgattgaa caagatggat
     3961 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4021 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4081 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4141 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4201 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4261 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4321 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4381 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4441 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4501 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4561 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4621 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4681 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4741 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca
     4801 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga
     4861 taaggatccg cgtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4921 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4981 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5041 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5101 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5161 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5221 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5281 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5341 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5401 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5461 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5521 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5581 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5641 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5701 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5761 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5821 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5881 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5941 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6001 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6061 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6121 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6181 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6241 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6301 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6361 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6421 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6481 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6541 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6601 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6661 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6721 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6781 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6841 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6901 tacggttcct ggccttttgc tggccttttg ctcacatggc tcgacagatc t
//
