LOCUS       sequence-290888-        7216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAGO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7216)
  AUTHORS   Kuzuoglu-Ozturk D, Bhandari D, Huntzinger E, Fauser M, Helms S, 
            Izaurralde E
  TITLE     miRISC and the CCR4-NOT complex silence mRNA targets independently 
            of 43S ribosomal scanning.
  JOURNAL   EMBO J. 2016 Jun 1;35(11):1186-203. doi: 10.15252/embj.201592901. 
            Epub 2016 Mar 23.
  PUBMED    27009120
REFERENCE   2  (bases 1 to 7216)
  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: 
            "10.15252/embj.201592901"; journalName: "EMBO J"; date: "2016-06-1- 
            1"; volume: "35"; issue: "11"; pages: "1186-203"
COMMENT     Sequence Label: pCMV-TAPtagN-PrP-SBP-HsAGO2_AC
FEATURES             Location/Qualifiers
     source          1..7216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      658..667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..867
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             868..1041
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1078..1101
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1108..1218
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     regulatory      3190..3199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4004..4125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4041..4060)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4095..4114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4132..4587)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4269..4290)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4481..4500
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4614..4718
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4720..5077
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4720..4911
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4743..4763)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4928..5063
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5112..5906
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5112..5906
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5166..5185)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5776..5795
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6094..6113)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6138..6185
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6514..7102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7003..7022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtgcc
      661 accatggcag gccttgcgca acacgatgaa gccgtggaca acaaattcaa caaagaacaa
      721 caaaacgcgt tctatgagat cttacattta cctaacttaa acgaagaaca acgaaacgcc
      781 ttcatccaaa gtttaaaaga tgacccaagc caaagcgcta accttttagc agaagctaaa
      841 aagctaaatg atgctcaggc gccgaaagta gacaacaaat tcaacaaaga acaacaaaac
      901 gcgttctatg agatcttaca tttacctaac ttaaacgaag aacaacgaaa cgccttcatc
      961 caaagtttaa aagatgaccc aagccaaagc gctaaccttt tagcagaagc taaaaagcta
     1021 aatgatgctc aggcgccgaa agtagacgcg aattgtgata tacctacaac tgcttctctg
     1081 gaagttctgt tccaggggcc cgaactagac gagaagacca ccggctggcg gggcggccac
     1141 gtggtggagg gcctggccgg cgagctggag cagctgcggg ccaggctgga gcaccaccct
     1201 cagggccagc gggagcccga tccatccgga ctcagatctc gagctcaagc ttatatggga
     1261 gggcacccat tccagtggtg taacgcactt gcacctcctg cgccgccgcc ccccatccaa
     1321 ggatatgcct tcaagcctcc acctagaccc gactttggga cctccgggag aacaatcaaa
     1381 ttacaggcca atttcttcga aatggacatc cccaaaattg acatctatca ttatgaattg
     1441 gatatcaagc cagagaagtg cccgaggaga gttaacaggg aaatcgtgga acacatggtc
     1501 cagcacttta aaacacagat ctttggggat cggaagcccg tgtttgacgg caggaagaat
     1561 ctatacacag ccatgcccct tccgattggg agggacaagg tggagctgga ggtcacgctg
     1621 ccaggagaag gcaaggatcg catcttcaag gtgtccatca agtgggtgtc ctgcgtgagc
     1681 ttgcaggcgt tacacgatgc actttcaggg cggctgccca gcgtcccttt tgagacgatc
     1741 caggccctgg acgtggtcat gaggcacttg ccatccatga ggtacacccc cgtgggccgc
     1801 tccttcttca ccgcgtccga aggctgctct aaccctcttg gcgggggccg agaagtgtgg
     1861 tttggcttcc atcagtccgt ccggccttct ctctggaaaa tgatgctgaa tattgatgtg
     1921 tcagcaacag cgttttacaa ggcacagcca gtaatcgagt ttgtttgtga agttttggat
     1981 tttaaaagta ttgaagaaca acaaaaacct ctgacagatt cccaaagggt aaagtttacc
     2041 aaagaaatta aaggtctaaa ggtggagata acgcactgtg ggcagatgaa gaggaagtac
     2101 cgtgtctgca atgtgacccg gcggcccgcc agtcaccaaa cattcccgct gcagcaggag
     2161 agcgggcaga cggtggagtg cacggtggcc cagtatttca aggacaggca caagttggtt
     2221 ctgcgctacc cccacctccc atgtttacaa gtcggacagg agcagaaaca cacctacctt
     2281 cccctggagg tctgtaacat tgtggcagga caaagatgta ttaaaaaatt aacggacaat
     2341 cagacctcaa ccatgatcag agcgactgct aggtcggcgc ccgatcggca agaagagatt
     2401 agcaaattga tgcgaagtgc aagtttcaac acagatccat acgtccgtga atttggaatc
     2461 atggtcaaag atgagatgac agacgtgact gggcgggtgc tgcagccgcc ctccatcctc
     2521 tacgggggca ggaataaagc tattgcgacc cctgtccagg gcgtctggga catgcggaac
     2581 aagcagttcc acacgggcat cgagatcaag gtgtgggcca ttgcgtgctt cgccccccag
     2641 cgccagtgca cggaagtcca tctgaagtcc ttcacagagc agctcagaaa gatctcgaga
     2701 gacgccggca tgcccatcca gggccagccg tgcttctgca aatacgcgca gggggcggac
     2761 agcgtggagc ccatgttccg gcacctgaag aacacgtatg cgggcctgca gctggtggtg
     2821 gtcatcctgc ccggcaagac gcccgtgtac gccgaggtca agcgcgtggg agacacggtg
     2881 ctggggatgg ccacgcagtg cgtgcagatg aagaacgtgc agaggaccac gccacagacc
     2941 ctgtccaacc tctgcctgaa gatcaacgtc aagctgggag gcgtgaacaa catcctgctg
     3001 ccccagggca ggccgccggt gttccagcag cccgtcatct ttctgggagc agacgtcact
     3061 cacccccccg ccggggatgg gaagaagccc tccattgccg ccgtggtggg cagcatggac
     3121 gcccacccca atcgctactg cgccaccgtg cgcgtgcagc agcaccggca ggagatcata
     3181 caagacctgg ccgccatggt ccgcgagctc ctcatccagt tctacaagtc cacgcgcttc
     3241 aagcccaccc gcatcatctt ctaccgcgac ggtgtctctg aaggccagtt ccagcaggtt
     3301 ctccaccacg agttgctggc catccgtgag gcctgtatca agctagaaaa agactaccag
     3361 cccgggatca ccttcatcgt ggtgcagaag aggcaccaca cccggctctt ctgcactgac
     3421 aagaacgagc gggttgggaa aagtggaaac attccagcag gcacgactgt ggacacgaaa
     3481 atcacccacc ccaccgagtt cgacttctac ctgtgtagtc acgctggcat ccaggggaca
     3541 agcaggcctt cgcactatca cgtcctctgg gacgacaatc gtttctcctc tgatgagctg
     3601 cagatcctaa cctaccagct gtgtcacacc tacgtgcgct gcacacgctc cgtgtccatc
     3661 ccagcgccag catactacgc tcacctggtg gccttccggg ccaggtacca cctggtggat
     3721 aaggaacatg acagtgctga aggaagccat acctctgggc agagtaacgg gcgagaccac
     3781 caagcactgg ccaaggcggt ccaggttcac caagacactc tgcgcaccat gtactttgct
     3841 tgagaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat
     3901 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3961 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4021 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4081 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     4141 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4201 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4261 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4321 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4381 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4441 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4501 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4561 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4621 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4681 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4741 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4801 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4861 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4921 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4981 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5041 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     5101 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5161 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5221 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5281 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5341 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5401 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5461 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5521 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5581 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5641 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5701 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5761 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5821 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5881 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5941 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6001 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6061 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     6121 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     6181 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     6241 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6301 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     6361 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6421 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6661 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7141 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7201 tattaccgcc atgcat
//
