LOCUS       sequence-106493-        8408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-Ago2 (N-termi tag) mutated five serines to alanines 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8408)
  AUTHORS   Horman SR, Janas MM, Litterst C, Wang B, MacRae IJ, Sever MJ, 
            Morrissey DV, Graves P, Luo B, Umesalma S, Qi HH, Miraglia LJ, 
            Novina CD, Orth AP
  TITLE     Akt-mediated phosphorylation of argonaute 2 downregulates cleavage 
            and upregulates translational repression of MicroRNA targets.
  JOURNAL   Mol Cell. 2013 May 9;50(3):356-67. doi: 
            10.1016/j.molcel.2013.03.015. Epub 2013 Apr 18.
  PUBMED    23603119
REFERENCE   2  (bases 1 to 8408)
  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: 
            "10.1016/j.molcel.2013.03.015"; journalName: "Mol Cell"; date: 
            "2013-05-9- 9"; volume: "50"; issue: "3"; pages: "356-67"
COMMENT     Sequence Label: pDEST26 Ago2 5A
FEATURES             Location/Qualifiers
     source          1..8408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(47..65)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(76..93)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(76..92)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(85..107)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    497..631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(534..553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     588..607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      757..1212
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(844..863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1054..1075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1331..1688
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1331..1522
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1354..1374)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1539..1674
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1739..2533
                     /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(1793..1812)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2403..2422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2597..2645
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2686..2705)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2805..2827
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2865..2883)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2951..3055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3056..3124,3125..3916)
                     /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(3274..3293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4087..4675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4576..4595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4829..4846
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        5135..5438
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5439..5642
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5593..5613
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5639..5663
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5764..5781
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    5791..5815
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      7744..7753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(join(8398..8408,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggttgatcg cgtgcatgcg acgtcatagc tctctcccta tagtgagtcg
       61 tattataagc taggcactgg ccgtcgtttt acaacgtcgt gactgggaaa actgctagct
      121 tgggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac
      181 agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactagc
      241 tgcatatgct tgctgcttga gagttttgct tactgagtat gatttatgaa aatattatac
      301 acaggagcta gtgattctaa ttgtttgtgt attttagatt cacagtccca aggctcattt
      361 caggcccctc agtcctcaca gtctgttcat gatcataatc agccatacca catttgtaga
      421 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
      481 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
      541 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
      601 actcatcaat gtatcttatc atgtctggat cgatcctgca ttaatgaatc ggccaacgcg
      661 cggggagagg cggtttgcgt attggctggc gtaatagcga agaggcccgc accgatcgcc
      721 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
      781 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
      841 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
      901 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
      961 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1021 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1081 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1141 attggttaaa aaatgagctg atttaacaaa tatttaacgc gaattttaac aaaatattaa
     1201 cgtttacaat ttcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     1261 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     1321 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     1381 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1441 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1501 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1561 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     1621 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     1681 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat
     1741 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     1801 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     1861 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     1921 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     1981 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2041 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2101 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2161 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     2221 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     2281 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     2341 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     2401 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     2461 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     2521 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     2581 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     2641 gtgtgaatcg atagcgataa ggatccgcgt atggtgcact ctcagtacaa tctgctctga
     2701 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2761 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2821 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     2881 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2941 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3001 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3061 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3121 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3181 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3241 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3301 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3361 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3421 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3481 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3541 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3601 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3661 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3721 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3781 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3841 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3901 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3961 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4021 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4081 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4141 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4201 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     4261 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4321 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4381 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4441 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     4501 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4561 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4621 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4681 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4741 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4801 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4861 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gagcttgcaa
     4921 ttcgcgcgtt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4981 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5041 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5101 tagtacgagg ccctttcact cattagatgc atgtcgttac ataacttacg gtaaatggcc
     5161 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     5221 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     5281 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     5341 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     5401 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     5461 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     5521 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5581 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5641 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     5701 gaagacaccg ggaccgatcc agcctccgga ctctagccta ggccgcggac catggcgtac
     5761 taccatcacc atcaccatca ctctagatca acaagtttgt acaaaaaagc aggcttaatg
     5821 tactcgggag ccggccccgc acttgcacct cctgcgccgc cgccccccat ccaaggatat
     5881 gccttcaagc ctccacctag acccgacttt gggacctccg ggagaacaat caaattacag
     5941 gccaatttct tcgaaatgga catccccaaa attgacatct atcattatga attggatatc
     6001 aagccagaga agtgcccgag gagagttaac agggaaatcg tggaacacat ggtccagcac
     6061 tttaaaacac agatctttgg ggatcggaag cccgtgtttg acggcaggaa gaatctatac
     6121 acagccatgc cccttccgat tgggagggac aaggtggagc tggaggtcac gctgccagga
     6181 gaaggcaagg atcgcatctt caaggtgtcc atcaagtggg tgtcctgcgt gagcttgcag
     6241 gcgttacacg atgcactttc agggcggctg cccagcgtcc cttttgagac gatccaggcc
     6301 ctggacgtgg tcatgaggca cttgccatcc atgaggtaca cccccgtggg ccgctccttc
     6361 ttcaccgcgt ccgaaggctg ctctaaccct cttggcgggg gccgagaagt gtggtttggc
     6421 ttccatcagt ccgtccggcc ttctctctgg aaaatgatgc tgaatattga tgtgtcagca
     6481 acagcgtttt acaaggcaca gccagtaatc gagtttgttt gtgaagtttt ggattttaaa
     6541 agtattgaag aacaacaaaa acctctgaca gattcccaaa gggtaaagtt taccaaagaa
     6601 attaaaggtc taaaggtgga gataacgcac tgtgggcaga tgaagaggaa gtaccgcgtc
     6661 tgcaatgtga cccggcggcc cgccagtcac caaacattcc cgctgcagca ggagagcggg
     6721 cagacggtgg agtgcacggt ggcccagtat ttcaaggaca ggcacaagtt ggttctgcgc
     6781 tacccccacc tcccatgttt acaagtcgga caggagcaga aacacaccta ccttcccctg
     6841 gaggtctgta acattgtggc aggacaaaga tgtattaaaa aattaacgga caatcagacc
     6901 tcaaccatga tcagagcgac tgctaggtcg gcgcccgatc ggcaagaaga gattagcaaa
     6961 ttgatgcgaa gtgcagcttt caacacagat ccatacgtcc gtgaatttgg aatcatggtc
     7021 aaagatgaga tgacagacgt gactgggcgg gtgctgcagc cgccctccat cctctacggg
     7081 ggcaggaata aagctattgc gacccctgtc cagggcgtct gggacatgcg gaacaagcag
     7141 ttccacacgg gcatcgagat caaggtgtgg gccattgcgt gcttcgcccc ccagcgccag
     7201 tgcacggaag tccatctgaa gtccttcaca gagcagctca gaaagatctc gagagacgcc
     7261 ggcatgccca tccagggcca gccgtgcttc tgcaaatacg cgcagggggc ggacagcgtg
     7321 gagcccatgt tccggcacct gaagaacacg tatgcgggcc tgcagctggt ggtggtcatc
     7381 ctgcccggca agacgcccgt gtacgccgag gtcaagcgcg tgggagacac ggtgctgggg
     7441 atggccacgc agtgcgtgca gatgaagaac gtgcagagga ccacgccaca gaccctgtcc
     7501 aacctctgcc tgaagatcaa cgtcaagctg ggaggcgtga acaacatcct gctgccccag
     7561 ggcaggccgc cggtgttcca gcagcccgtc atctttctgg gagcagacgt cactcacccc
     7621 cccgccgggg atgggaagaa gccctccatt gccgccgtgg tgggcagcat ggacgcccac
     7681 cccaatcgct actgcgccac cgtgcgtgtg cagcagcacc ggcaggagat catacaagac
     7741 ctggccgcca tggtccgcga gctcctcatc cagttctaca agtccacgcg cttcaagccc
     7801 acccgcatca tcttctaccg cgacggtgtc tctgaaggcc agttccagca ggttctccac
     7861 cacgagttgc tggccatccg tgaggcctgt atcaagctag aaaaagacta ccagccaggg
     7921 atcaccttca tcgtggtgca gaagaggcac cacacccggc tcttctgcac tgacaagaac
     7981 gagcgggttg ggaaaagtgg aaacattcca gcaggcacga ctgtggacac gaaaatcacc
     8041 caccccaccg agttcgactt ctacctgtgt agtcacgctg gcatccaggg gacaagcagg
     8101 ccttcgcact atcacgtcct ctgggacgac aatcgtttct cctctgatga gctgcagatc
     8161 ctaacctacc agctgtgtca cacctacgtg cgctgcacac gctccgtgtc catcccagcg
     8221 ccagcatact acgctcacct ggtggccttc cgggccaggt accacctggt ggataaggaa
     8281 catgacgctg ctgaaggagc ccataccgct gggcaggcta acgggcgaga ccaccaagca
     8341 ctggccaagg cggtccaggt tcaccaagac actctgcgca ccatgtactt tgcttgaacc
     8401 cagctttc
//
