LOCUS       sequence-430706-        7844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AGO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7844)
  AUTHORS   Nishi K, Takahashi T, Suzawa M, Miyakawa T, Nagasawa T, Ming Y, 
            Tanokura M, Ui-Tei K
  TITLE     Control of the localization and function of a miRNA silencing 
            component TNRC6A by Argonaute protein.
  JOURNAL   Nucleic Acids Res. 2015 Nov 16;43(20):9856-73. doi: 
            10.1093/nar/gkv1026. Epub 2015 Oct 7.
  PUBMED    26446993
REFERENCE   2  (bases 1 to 7844)
  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: 
            "10.1093/nar/gkv1026"; journalName: "Nucleic Acids Res"; date: 
            "2015-11-16- 16"; volume: "43"; issue: "20"; pages: "9856-73"
COMMENT     Sequence Label: pIRESneo-Ago2
FEATURES             Location/Qualifiers
     source          1..7844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2853..2862
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3585..3814
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3797..3816
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3887..4461,4462..4464,4465..4473)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4054..4071)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4281..4300
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4496..5299
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4559..4578)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5169..5188
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5331..5348)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5337..5561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5608..5627
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5687..5703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5687..5703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5700..5722)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5711..5727
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5735..5741,5742..5759,5760..5765))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5780..5801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5918..5935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6089..6677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6169..6188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6848..7639,7640..7708))
                     /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     7471..7490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7709..7813)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatcgcc gccgccatgt actcgggagc cggccccgca cttgcacctc
      961 ctgcgccgcc gccccccatc caaggatatg ccttcaagcc tccacctaga cccgactttg
     1021 ggacctccgg gagaacaatc aaattacagg ccaatttctt cgaaatggac atccccaaaa
     1081 ttgacatcta tcattatgaa ttggatatca agccagagaa gtgcccgagg agagttaaca
     1141 gggaaatcgt ggaacacatg gtccagcact ttaaaacaca gatctttggg gatcggaagc
     1201 ccgtgtttga cggcaggaag aatctataca cagccatgcc ccttccgatt gggagggaca
     1261 aggtggagct ggaggtcacg ctgccaggag aaggcaagga tcgcatcttc aaggtgtcca
     1321 tcaagtgggt gtcctgcgtg agcttgcagg cgttacacga tgcactttca gggcggctgc
     1381 ccagcgtccc ttttgagacg atccaggccc tggacgtggt catgaggcac ttgccatcca
     1441 tgaggtacac ccccgtgggc cgctccttct tcaccgcgtc cgaaggctgc tctaaccctc
     1501 ttggcggggg ccgagaagtg tggtttggct tccatcagtc cgtccggcct tctctctgga
     1561 aaatgatgct gaatattgat gtgtcagcaa cagcgtttta caaggcacag ccagtaatcg
     1621 agtttgtttg tgaagttttg gattttaaaa gtattgaaga acaacaaaaa cctctgacag
     1681 attcccaaag ggtaaagttt accaaagaaa ttaaaggtct aaaggtggag ataacgcact
     1741 gtgggcagat gaagaggaag taccgtgtct gcaatgtgac ccggcggccc gccagtcacc
     1801 aaacattccc gctgcagcag gagagcgggc agacggtgga gtgcacggtg gcccagtatt
     1861 tcaaggacag gcacaagttg gttctgcgct acccccacct cccatgttta caagtcggac
     1921 aggagcagaa acacacctac cttcccctgg aggtctgtaa cattgtggca ggacaaagat
     1981 gtattaaaaa attaacggac aatcagacct caaccatgat cagagcgact gctaggtcgg
     2041 cgcccgatcg gcaagaagag attagcaaat tgatgcgaag tgcaagtttc aacacagatc
     2101 catacgtccg tgaatttgga atcatggtca aagatgagat gacagacgtg actgggcggg
     2161 tgctgcagcc gccctccatc ctctacgggg gcaggaataa agctattgcg acccctgtcc
     2221 agggcgtctg ggacatgcgg aacaagcagt tccacacggg catcgagatc aaggtgtggg
     2281 ccattgcgtg cttcgccccc cagcgccagt gcacggaagt ccatctgaag tccttcacag
     2341 agcagctcag aaagatctcg agagacgccg gcatgcccat ccagggccag ccgtgcttct
     2401 gcaaatacgc gcagggggcg gacagcgtgg agcccatgtt ccggcacctg aagaacacgt
     2461 atgcgggcct gcagctggtg gtggtcatcc tgcccggcaa gacgcccgtg tacgccgagg
     2521 tcaagcgcgt gggagacacg gtgctgggga tggccacgca gtgcgtgcag atgaagaacg
     2581 tgcagaggac cacgccacag accctgtcca acctctgcct gaagatcaac gtcaagctgg
     2641 gaggcgtgaa caacatcctg ctgccccagg gcaggccgcc ggtgttccag cagcccgtca
     2701 tctttctggg agcagacgtc actcaccccc ccgccgggga tgggaagaag ccctccattg
     2761 ccgccgtggt gggcagcatg gacgcccacc ccaatcgcta ctgcgccacc gtgcgcgtgc
     2821 agcagcaccg gcaggagatc atacaagacc tggccgccat ggtccgcgag ctcctcatcc
     2881 agttctacaa gtccacgcgc ttcaagccca cccgcatcat cttctaccgc gacggtgtct
     2941 ctgaaggcca gttccagcag gttctccacc acgagttgct ggccatccgt gaggcctgta
     3001 tcaagctaga aaaagactac cagcccggga tcaccttcat cgtggtgcag aagaggcacc
     3061 acacccggct cttctgcact gacaagaacg agcgggttgg gaaaagtgga aacattccag
     3121 caggcacgac tgtggacacg aaaatcaccc accccaccga gttcgacttc tacctgtgta
     3181 gtcacgctgg catccagggg acaagcaggc cttcgcacta tcacgtcctc tgggacgaca
     3241 atcgtttctc ctctgatgag ctgcagatcc taacctacca gctgtgtcac acctacgtgc
     3301 gctgcacacg ctccgtgtcc atcccagcgc cagcatacta cgctcacctg gtggccttcc
     3361 gggccaggta ccacctggtg gataaggaac atgacagtgc tgaaggaagc catacctctg
     3421 ggcagagtaa cgggcgagac caccaagcac tggccaaggc ggtccaggtt caccaagaca
     3481 ctctgcgcac catgtacttt gcttgacgga attcagtgga tccactagta acggccgcca
     3541 gtgtgctgga attaattcgc tgtctgcgag ggccagctgt tggggtgagt actccctctc
     3601 aaaagcgggc atgacttctg cgctaagatt gtcagtttcc aaaaacgagg aggatttgat
     3661 attcacctgg cccgcggtga tgcctttgag ggtggccgcg tccatctggt cagaaaagac
     3721 aatctttttg ttgtcaagct tgaggtgtgg caggcttgag atctggccat acacttgagt
     3781 gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcag
     3841 gtcgagcatg catctagggc ggccaattcc gcccctctcc cccccccccc tctccctccc
     3901 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     3961 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     4021 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     4081 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     4141 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     4201 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     4261 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     4321 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     4381 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     4441 tttgaaaaac acgatgataa gcttgccaca acccgggata attcctgcag ccaatatggg
     4501 atcggccatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4561 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4621 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4681 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4741 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4801 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4861 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4921 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4981 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5041 cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5101 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5161 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5221 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5281 tcttgacgag ttcttctgag gggatcaatt ctctagagct cgctgatcag cctcgactgt
     5341 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5401 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5461 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5521 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     5581 cagctggggc tcgagtgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5641 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5701 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5761 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5821 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5881 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6961 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7141 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7681 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7741 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7801 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
