LOCUS       sequence-289036-        6655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6C_1248-1600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  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 6655)
  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_1248-1600_I
FEATURES             Location/Qualifiers
     source          1..6655
                     /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    2359..2480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2370..2389)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2424..2443
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2666..3121
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2753..2772)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2963..2984
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3238..3595
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3238..3429
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3261..3281)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3446..3581
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3646..4440
                     /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(3700..3719)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4310..4329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4504..4552
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4593..4612)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4712..4734
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4772..4790)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4858..4962
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4963..5031,5032..5823)
                     /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(5181..5200)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5994..6582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6483..6502
                     /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 acctcaggaa
     2041 gaaccagcag ctggctcgtt cttcgaaacc tcactcccca gattgatggt tctacactgc
     2101 ggacattgtg tttgcaacat gggcctctta tcacattcca cctgaatctg actcaaggca
     2161 atgctgtggt ccggtacagc tccaaggagg aggctgccaa ggcccagaag tctctgcaca
     2221 tgtgcgtcct gggaaacact accatcctgg ccgagttcgc tggtgaagaa gaagtgaatc
     2281 gcttcttagc ccaaggccag gcgctgccat aggcggccgc ttccctttag tgagggttaa
     2341 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2401 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2461 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2521 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     2581 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2641 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2701 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2761 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2821 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2881 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2941 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3001 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3061 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3121 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     3181 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     3241 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3301 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3361 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3421 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3481 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3541 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3601 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     3661 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3721 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3781 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3841 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3901 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3961 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4021 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4081 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4141 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4201 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4261 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4321 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4381 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4441 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     4501 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     4561 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4621 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4681 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4741 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4801 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4861 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4921 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4981 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5041 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5101 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5161 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5221 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5281 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5341 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5401 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5461 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5521 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5581 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5641 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5701 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5761 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5821 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5881 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5941 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6121 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6241 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6301 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6361 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6421 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6541 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6601 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatct
//
