LOCUS       sequence-333971-        8147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAGO3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8147)
  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 8147)
  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-HsAGO3_C
FEATURES             Location/Qualifiers
     source          1..8147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      952..961
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             958..1023
                     /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             1042..1068
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3714..3835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3725..3744)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3779..3798
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4021..4476
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4108..4127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4318..4339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4593..4950
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4593..4784
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4616..4636)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4801..4936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5001..5795
                     /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(5055..5074)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5665..5684
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5859..5907
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5948..5967)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6067..6089
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6127..6145)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6213..6317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6318..6386,6387..7178)
                     /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(6536..6555)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7349..7937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7838..7857
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agccaccatg
      961 gacgcacaaa cacgacgacg tgagcgtcgc gctgagaaac aagctcaatg gaaagctgca
     1021 aacccaccgc tcgagatggc ttacccatac gatgttccag attacgctat ggaaatcggc
     1081 tccgcaggac ccgctggggc ccagccccta ctcatggtgc ccagaagacc tggctatggc
     1141 gccatgggca aacccattaa actgctggct aactgttttc aagttgaaat cccaaagatt
     1201 gatgtctacc tctatgaggt agatattaaa ccagacaagt gtcctaggag agtgaacagg
     1261 gaggtggttg actcaatggt tcagcatttt aaagtaacta tatttggaga ccgtagacca
     1321 gtttatgatg gaaaaagaag tctttacacc gccaatccac ttcctgtggc aactacaggg
     1381 gtagatttag acgttacttt acctggggaa ggtggaaaag atcgaccttt caaggtgtca
     1441 atcaaatttg tctctcgggt gagttggcac ctactgcatg aagtactgac aggacggact
     1501 ttgcctgagc cactggaatt agacaagcca atcagcacta accctgtcca tgccgttgat
     1561 gtggtgctac gacatctgcc ctccatgaaa tacacacctg tggggcgttc atttttctcc
     1621 gctccagaag gatatgacca ccctctggga gggggcaggg aagtgtggtt tggattccat
     1681 cagtctgttc ggcctgccat gtggaaaatg atgcttaata tcgatgtttc tgccactgcc
     1741 ttctacaaag cacaacctgt aattcagttc atgtgtgaag ttcttgatat tcataatatt
     1801 gatgagcaac caagacctct gactgattct catcgggtaa aattcaccaa agagataaaa
     1861 ggtttgaagg ttgaagtgac tcattgtgga acaatgagac ggaaataccg tgtttgtaat
     1921 gtaacaagga ggcctgccag tcatcaaacc tttcctttac agttagaaaa cggccaaact
     1981 gtggagagaa cagtagcgca gtatttcaga gaaaagtata ctcttcagct gaagtacccg
     2041 caccttccct gtctgcaagt cgggcaggaa cagaaacaca cctacctgcc actagaagtc
     2101 tgtaatattg tggcagggca acgatgtatc aagaagctaa cagacaatca gacttccact
     2161 atgatcaagg caacagcaag atctgcacca gatagacaag aggaaattag cagattggta
     2221 agaagtgcaa attatgaaac agatccattt gttcaggagt ttcaatttaa agttcgggat
     2281 gaaatggctc atgtaactgg acgcgtactt ccagcaccta tgctccagta tggaggacgg
     2341 aatcggacag tagcaacacc gagccatgga gtatgggaca tgcgagggaa acaattccac
     2401 acaggagttg aaatcaaaat gtgggctatc gcttgttttg ccacacagag gcagtgcaga
     2461 gaagaaatat tgaagggttt cacagaccag ctgcgtaaga tttctaagga tgcagggatg
     2521 cccatccagg gccagccatg cttctgcaaa tatgcacagg gggcagacag cgtagagccc
     2581 atgttccggc atctcaagaa cacatattct ggcctacagc ttattatcgt catcctgccg
     2641 gggaagacac cagtgtatgc ggaagtgaaa cgtgtaggag acacactttt gggtatggct
     2701 acacaatgtg ttcaagtcaa gaatgtaata aaaacatctc ctcaaactct gtcaaacttg
     2761 tgcctaaaga taaatgttaa actcggaggg atcaataata ttcttgtacc tcatcaaaga
     2821 ccttctgtgt tccagcaacc agtgatcttt ttgggagccg atgtcactca tccacctgct
     2881 ggtgatggaa agaagccttc tattgctgct gttgtaggta gtatggatgc acacccaagc
     2941 agatactgtg ccacagtaag agttcagaga ccccgacagg agatcatcca ggacttggcc
     3001 tccatggtcc gggaacttct tattcaattt tataagtcaa ctcggttcaa gcctactcgt
     3061 atcatctttt atcgggatgg tgtttcagag gggcagttta ggcaggtatt atattatgaa
     3121 ctactagcaa ttcgagaagc ctgcatcagt ttggagaaag actatcaacc tggaataacc
     3181 tacattgtag ttcagaagag acatcacact cgattatttt gtgctgatag gacagaaagg
     3241 gttggaagaa gtggcaatat cccagctgga acaacagttg atacagacat tacacaccca
     3301 tatgagttcg atttttacct ctgtagccat gctggaatac agggtaccag tcgtccttca
     3361 cactatcatg ttttatggga tgataactgc tttactgcag atgaacttca gctgctaact
     3421 taccagctct gccacactta cgtacgctgt acacgatctg tttctatacc tgcaccagcg
     3481 tattatgctc acctggtagc atttagagcc agatatcatc ttgtggacaa agaacatgac
     3541 agtgctgaag gaagtcacgt ttcaggacaa agcaatgggc gagatccaca agctcttgcc
     3601 aaggctgtac agattcacca agatacctta cgcacaatgt acttcgctta atctagagtc
     3661 gacccgggcg gccgcttccc tttagtgagg gttaatgctt cgagcagaca tgataagata
     3721 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3781 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3841 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     3901 caagtaaaac ctctacaaat gtggtaaaat ccgataagga tcgatccggg ctggcgtaat
     3961 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4021 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4081 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4141 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4201 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4261 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4321 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4381 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4441 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     4501 gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata
     4561 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     4621 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4681 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4741 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4801 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4861 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4921 ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg
     4981 aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5041 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5101 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5161 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5221 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5281 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5341 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5401 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5461 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5521 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5581 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5641 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5701 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5761 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5821 cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat
     5881 tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca
     5941 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     6001 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6061 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     6121 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6181 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6241 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6301 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6361 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6421 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6481 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6541 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6601 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6661 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6721 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6781 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6841 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6901 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6961 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7021 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7081 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7141 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7201 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7261 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7321 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7381 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7441 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7501 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7561 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7621 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7681 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7741 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7801 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7861 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7921 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7981 ggccttttgc tcacatggct cgacagatct tcaatattgg ccattagcca tattattcat
     8041 tggttatata gcataaatca atattggcta ttggccattg catacgttgt atctatatca
     8101 taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgtt
//
