LOCUS       sequence-288530-        8146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAGO4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8146)
  AUTHORS   Lazzaretti D, Tournier I, Izaurralde E
  TITLE     The C-terminal domains of human TNRC6A, TNRC6B, and TNRC6C silence 
            bound transcripts independently of Argonaute proteins.
  JOURNAL   RNA. 2009 Jun . 15(6):1059-66.
  PUBMED    19383768
REFERENCE   2  (bases 1 to 8146)
  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"; journalName: "RNA. 2009 
            Jun . 15(6):1059-66."
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsAGO4_C
FEATURES             Location/Qualifiers
     source          1..8146
                     /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             1191..1217
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3850..3971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3861..3880)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3915..3934
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4157..4612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4244..4263)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4454..4475
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4729..5086
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4729..4920
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4752..4772)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4937..5072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5137..5931
                     /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(5191..5210)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5801..5820
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5995..6043
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6084..6103)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6203..6225
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6263..6281)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6349..6453
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6454..6522,6523..7314)
                     /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(6672..6691)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7485..8073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7974..7993
                     /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 agaattcacg cgtgatggct tacccatacg
     1201 atgttccaga ttacgctatg gaggcgctgg gacccggacc tccggctagc ctgtttcagc
     1261 cacctcgtcg tcctggcctt ggaactgttg gaaaaccaat tcgactgtta gccaatcatt
     1321 ttcaggttca gattcctaaa atagatgtgt atcactatga tgtggatatt aagcctgaaa
     1381 aacggcctcg tagagtcaac agggaggtag tagatacaat ggtgcggcac ttcaagatgc
     1441 aaatatttgg tgatcggcag cctgggtatg atggcaaaag aaacatgtac acagcacatc
     1501 cactaccaat tggacgggat agggttgata tggaggtgac tcttccaggc gagggtaaag
     1561 accaaacatt taaagtgtct gttcagtggg tgtcagttgt gagccttcag ttgcttttag
     1621 aagctttggc tgggcacttg aatgaagtcc cagatgactc agtacaagca cttgatgtta
     1681 tcacaagaca ccttccctcc atgaggtaca ccccagtggg ccgttccttt ttctcacccc
     1741 cggaaggtta ctaccaccct ctgggagggg gcagggaggt ctggtttggt tttcatcagt
     1801 ctgtgagacc tgccatgtgg aatatgatgc tcaacattga tgtatctgca actgctttct
     1861 accgggctca gcctatcatt gagttcatgt gtgaggtttt agacattcag aacatcaatg
     1921 aacagaccaa acctctaaca gactcccagc gtgtcaaatt taccaaagaa atcagaggtc
     1981 tcaaagttga ggtgacccac tgtggacaga tgaaacgaaa ataccgagtt tgtaatgtga
     2041 ctagacggcc agccagtcat caaacttttc ctttgcagct agaaaacggt caagctatgg
     2101 aatgtacagt agctcaatat tttaagcaaa agtatagtct gcaactgaaa tacccccatc
     2161 ttccctgtct ccaagtggga caagaacaaa agcatacata cttgccactc gaggtctgta
     2221 atatagtggc aggacagcga tgtatcaaga agctcacaga caatcagact tccacaatga
     2281 tcaaagctac agcaagatct gctcctgaca gacaggaaga gatcagtaga ctggtgaaga
     2341 gcaacagtat ggtgggtgga cctgatccat accttaaaga atttggtatt gttgtccaca
     2401 atgaaatgac agagctcaca ggcagggtac ttccagcacc aatgctgcaa tatggaggcc
     2461 ggaataaaac agtagccaca cccaaccagg gtgtctggga catgcgagga aagcagtttt
     2521 atgctggcat tgaaattaaa gtttgggcag ttgcttgttt tgcacctcag aaacaatgta
     2581 gggaagattt actaaagagt ttcactgacc agctgcgtaa aatctctaag gatgcaggaa
     2641 tgcccatcca gggtcagcca tgtttctgca agtatgcaca aggtgcagac agtgtggagc
     2701 ctatgtttaa acatctgaaa atgacttatg tgggcctaca gctaatagtg gttatcctgc
     2761 ctggaaagac accagtatat gcggaggtga aacgtgttgg agataccctt ctaggtatgg
     2821 ccacacagtg tgtccaggta aaaaatgtag tgaagacctc acctcaaacc ctttccaatc
     2881 tttgcctgaa gataaatgca aaacttggag gaattaacaa tgtgcttgtg cctcatcaaa
     2941 ggccctcggt gttccagcag cctgtcatct tcctgggagc ggatgtcaca caccccccag
     3001 caggggatgg gaagaaacct tccattgctg ctgtggttgg cagtatggat ggccacccca
     3061 gccggtactg tgccaccgtt cgggtgcaga cttcccggca ggagatctcc caagagctcc
     3121 tctacagtca agaggtcatc caggacctga ctaacatggt tcgagagctg ctgattcagt
     3181 tctacaaatc cacacgcttc aaacccactc ggatcatcta ttaccgtgga ggggtatctg
     3241 agggacaaat gaaacaggta gcttggccag aactaatagc aattcgaaag gcatgtatta
     3301 gcttggaaga agattaccgg ccaggaataa cttatattgt ggtgcaaaaa agacatcaca
     3361 cacgactctt ctgtgcagat aaaacagaaa gggtagggaa aagtggcaat gtaccagcag
     3421 gcactacagt ggatagtacc atcacacatc catctgagtt tgacttttac ctctgtagtc
     3481 atgcaggaat tcagggaacc agccgtccct cacattacca ggtcttgtgg gatgacaact
     3541 gcttcactgc agatgaactc cagctactga cttaccagct gtgtcacacc tatgtgaggt
     3601 gcactcgctc agtctctatt ccagcccctg catattatgc ccggcttgta gcatttaggg
     3661 caaggtatca tctggtggat aaagatcatg acagtgcgga aggcagtcat gtgtcaggac
     3721 agagcaacgg ccgggatcct caggccttgg ctaaggctgt gcaaatccac catgataccc
     3781 agcacacgat gtattttgcc tgagcggccg cttcccttta gtgagggtta atgcttcgag
     3841 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3901 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3961 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     4021 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatccga taaggatcga
     4081 tccgggctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4141 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4201 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4261 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4321 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4381 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4441 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4501 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4561 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     4621 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc
     4681 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     4741 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4801 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4861 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4921 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4981 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5041 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttgattcttc
     5101 tgacacaaca gtctcgaact taaggctaga gccaccatga ttgaacaaga tggattgcac
     5161 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5221 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5281 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5341 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5401 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5461 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5521 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5581 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5641 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5701 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5761 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5821 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5881 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5941 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa
     6001 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg
     6061 atccgcgtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     6121 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     6181 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     6241 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     6301 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     6361 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6421 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6481 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6541 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6601 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6661 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6721 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6781 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6841 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6901 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6961 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     7021 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7081 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7141 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7201 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     7261 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     7321 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7381 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7441 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7561 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7621 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7681 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7741 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7801 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7861 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7921 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7981 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8041 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8101 ttcctggcct tttgctggcc ttttgctcac atggctcgac agatct
//
