LOCUS       sequence-290875-        8900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAGO2-K663AL697Y.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8900)
  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 8900)
  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: pCIneo-lambdaN-HA-HsAGO2-K663AL697Y_AC
FEATURES             Location/Qualifiers
     source          1..8900
                     /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"
     regulatory      3189..3198
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4604..4725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4615..4634)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4669..4688
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4911..5366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4998..5017)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5208..5229
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5483..5840
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5483..5674
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5506..5526)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5691..5826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5891..6685
                     /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(5945..5964)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6555..6574
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6749..6797
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6838..6857)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6957..6979
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7017..7035)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7103..7207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7208..7276,7277..8068)
                     /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(7426..7445)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8239..8827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8728..8747
                     /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 accatgggag
     1261 ggcacccatt ccagtggtgt aacgcacttg cacctcctgc gccgccgccc cccatccaag
     1321 gatatgcctt caagcctcca cctagacccg actttgggac ctccgggaga acaatcaaat
     1381 tacaggccaa tttcttcgaa atggacatcc ccaaaattga catctatcat tatgaattgg
     1441 atatcaagcc agagaagtgc ccgaggagag ttaacaggga aatcgtggaa cacatggtcc
     1501 agcactttaa aacacagatc tttggggatc ggaagcccgt gtttgacggc aggaagaatc
     1561 tatacacagc catgcccctt ccgattggga gggacaaggt ggagctggag gtcacgctgc
     1621 caggagaagg caaggatcgc atcttcaagg tgtccatcaa gtgggtgtcc tgcgtgagct
     1681 tgcaggcgtt acacgatgca ctttcagggc ggctgcccag cgtccctttt gagacgatcc
     1741 aggccctgga cgtggtcatg aggcacttgc catccatgag gtacaccccc gtgggccgct
     1801 ccttcttcac cgcgtccgaa ggctgctcta accctcttgg cgggggccga gaagtgtggt
     1861 ttggcttcca tcagtccgtc cggccttctc tctggaaaat gatgctgaat attgatgtgt
     1921 cagcaacagc gttttacaag gcacagccag taatcgagtt tgtttgtgaa gttttggatt
     1981 ttaaaagtat tgaagaacaa caaaaacctc tgacagattc ccaaagggta aagtttacca
     2041 aagaaattaa aggtctaaag gtggagataa cgcactgtgg gcagatgaag aggaagtacc
     2101 gtgtctgcaa tgtgacccgg cggcccgcca gtcaccaaac attcccgctg cagcaggaga
     2161 gcgggcagac ggtggagtgc acggtggccc agtatttcaa ggacaggcac aagttggttc
     2221 tgcgctaccc ccacctccca tgtttacaag tcggacagga gcagaaacac acctaccttc
     2281 ccctggaggt ctgtaacatt gtggcaggac aaagatgtat taaaaaatta acggacaatc
     2341 agacctcaac catgatcaga gcaactgcta ggtcggcgcc cgatcggcaa gaagagatta
     2401 gcaaattgat gcgaagtgca agtttcaaca cagatccata cgtccgtgaa tttggaatca
     2461 tggtcaaaga tgagatgaca gacgtgactg ggcgggtgct gcagccgccc tccatcctct
     2521 acgggggcag gaataaagct attgcgaccc ctgtccaggg cgtctgggac atgcggaaca
     2581 agcagttcca cacgggcatc gagatcaagg tgtgggccat tgcgtgcttc gccccccagc
     2641 gccagtgcac ggaagtccat ctgaagtcct tcacagagca gctcagaaag atctcgagag
     2701 acgctggcat gcccatccag ggccagccgt gcttctgcaa atacgcgcag ggggcggaca
     2761 gcgtggagcc catgttccgg cacctgaaga acacgtatgc gggcctgcag ctggtggtgg
     2821 tcatcctgcc cggcaagacg cccgtgtacg ccgaggtcaa gcgcgtggga gacacggtgc
     2881 tggggatggc cacgcagtgc gtgcagatga agaacgtgca gaggaccacg ccacagaccc
     2941 tgtccaacct ctgcctgaag atcaacgtca agctgggagg cgtgaacaac atcctgctgc
     3001 cccagggcag gccgccggtg ttccagcagc ccgtcatctt tctgggagca gacgtcactc
     3061 acccccccgc cggggatggg aagaagccct ccattgccgc cgtggtgggc agcatggacg
     3121 cccaccccaa tcgctactgc gccaccgtgc gcgtgcagca gcaccggcag gagatcatac
     3181 aagacctggc cgccatggtc cgcgagctcc tcatccagtt ctacaagtcc acgcgcttcg
     3241 cgcccacccg catcatcttc taccgcgacg gtgtctctga aggccagttc cagcaggttc
     3301 tccaccacga gttgctggcc atccgtgagg cctgtatcaa gtatgaaaaa gactaccagc
     3361 ccgggatcac cttcatcgtg gtgcagaaga ggcaccacac ccggctcttc tgcactgaca
     3421 agaacgagcg ggttgggaaa agtggaaaca ttccagcagg cacgactgtg gacacgaaaa
     3481 tcacccaccc caccgagttc gacttctacc tgtgtagtca cgctggcatc caggggacaa
     3541 gcaggccttc gcactatcac gtcctctggg acgacaatcg tttctcctct gatgagctgc
     3601 agatcctaac ctaccagctg tgtcacacct acgtgcgctg cacacgctcc gtgtccatcc
     3661 cagcgccagc atactacgct cacctggtgg ccttccgggc caggtaccac ctggtggata
     3721 aggaacatga cagtgctgaa ggaagccata cctctgggca gagtaacggg cgagaccacc
     3781 aagcactggc caaggcggtc caggttcacc aagacactct gcgcaccatg tactttgctt
     3841 gacatgtttt agtgtttagc gattgtgtac cgagtgggat tcacgagacc agctacactc
     3901 agaccaacag atggccagcc cttccgtgac agccagcatc gaacatgaga cgtcattgat
     3961 tttattagat tctccgtttt ccagaatgcc ttccgtccca gatttcaaac ttggattttg
     4021 aactgcagac ctgtatgaga acccaatgtc ataggaaata tggtttgcta aaatctataa
     4081 gctgcttatt aaaacagagt cccgtgtgtc ctaaaaatct cctaaaacca gtctatgaac
     4141 tcagggcttt aaaacatttt taatttattt ggtcattcaa tttacttgtt tttaatacat
     4201 gattctctat gaaattgatg ggctcaaact agctgtgaat cttctgagag tgaaagcaac
     4261 acaaaacaca agtgtggttt taaagccttg aacattctga tgttgtcact aaagttgatt
     4321 tccaggcgat gctcgtgtgc ccctggcgtg gctcacccaa gttcctcgac tgagggccgg
     4381 tggccatgca gaggcgtccg caggtgccgt gttctgccag caccgccctt cacccggcct
     4441 gaactaagga gcagtggcag aaggtgggcc ccgtgtgttt acagcatttc caggtccaga
     4501 gaggttggca gacaagtgcc attttaataa aaaatgtatt taaaaaaaaa aaaaagggcg
     4561 gccgcttccc tttagtgagg gttaatgctt cgagcagaca tgataagata cattgatgag
     4621 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4681 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4741 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     4801 ctctacaaat gtggtaaaat ccgataagga tcgatccggg ctggcgtaat agcgaagagg
     4861 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     4921 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4981 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5041 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5101 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5161 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5221 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5281 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5341 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     5401 ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa
     5461 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     5521 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5581 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5641 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5701 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5761 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5821 gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaaggc
     5881 tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5941 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     6001 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     6061 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     6121 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     6181 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     6241 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     6301 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     6361 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     6421 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6481 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6541 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6601 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6661 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6721 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     6781 tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca ctctcagtac
     6841 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     6901 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     6961 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     7021 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     7081 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     7141 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     7201 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     7261 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     7321 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     7381 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     7441 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7501 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7561 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7621 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7681 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7741 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7801 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7861 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7921 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7981 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     8041 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     8101 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     8161 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8221 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     8281 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     8341 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     8401 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     8461 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8521 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8581 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8641 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8701 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8761 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8821 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8881 tcacatggct cgacagatct
//
