LOCUS       sequence-55293-e        7909 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7909)
  AUTHORS   Meister G, Landthaler M, Patkaniowska A, Dorsett Y, Teng G, Tuschl T
  TITLE     Human Argonaute2 mediates RNA cleavage targeted by miRNAs and 
            siRNAs.
  JOURNAL   Mol Cell. 2004 Jul 23. 15(2):185-97.
  PUBMED    15260970
REFERENCE   2  (bases 1 to 7909)
  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: "Mol Cell. 
            2004 Jul 23. 15(2):185-97."
COMMENT     Sequence Label: pIRESneo-FLAG/HA Ago2 corrected
FEATURES             Location/Qualifiers
     source          1..7909
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      1457..1466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          2189..2418
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2401..2420
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2491..3065
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2659..2676)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2886..2905
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3101..3904
                     /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(3164..3183)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3774..3793
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3936..3953)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3942..4166
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4213..4232
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4292..4308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4292..4308)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4305..4327)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4316..4332
                     /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(4340..4346,4347..4364,4365..4370))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4385..4406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4523..4540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4694..5282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4774..4793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5453..6244,6245..6313))
                     /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     6076..6095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6314..6418)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6493..6512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6684..7063
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7064..7267
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7218..7238
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7312..7331
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7312..7330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      7371..7380
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7380..7403
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7404..7430
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gacgatccag gccctggacg tggtcatgag gcacttgcca tccatgaggt acacccccgt
       61 gggccgctcc ttcttcaccg cgtccgaagg ctgctctaac cctcttggcg ggggccgaga
      121 agtgtggttt ggcttccatc agtccgtccg gccttctctc tggaaaatga tgctgaatat
      181 tgatgtgtca gcaacagcgt tttacaaggc acagccagta atcgagtttg tttgtgaagt
      241 tttggatttt aaaagtattg aagaacaaca aaaacctctg acagattccc aaagggtaaa
      301 gtttaccaaa gaaattaaag gtctaaaggt ggagataacg cactgtgggc agatgaagag
      361 gaagtaccgt gtctgcaatg tgacccggcg gcccgccagt caccaaacat tcccgctgca
      421 gcaggagagc gggcagacgg tggagtgcac ggtggcccag tatttcaagg acaggcacaa
      481 gttggttctg cgctaccccc acctcccatg tttacaagtc ggacaggagc agaaacacac
      541 ctaccttccc ctggaggtct gtaacattgt ggcaggacaa agatgtatta aaaaattaac
      601 ggacaatcag acctcaacca tgatcagagc gactgctagg tcggcgcccg atcggcaaga
      661 agagattagc aaattgatgc gaagtgcaag tttcaacaca gatccatacg tccgtgaatt
      721 tggaatcatg gtcaaagatg agatgacaga cgtgactggg cgggtgctgc agccgccctc
      781 catcctctac gggggcagga ataaagctat tgcgacccct gtccagggcg tctgggacat
      841 gcggaacaag cagttccaca cgggcatcga gatcaaggtg tgggccattg cgtgcttcgc
      901 cccccagcgc cagtgcacgg aagtccatct gaagtccttc acagagcagc tcagaaagat
      961 ctcgagagac gccggcatgc ccatccaggg ccagccgtgc ttctgcaaat acgcgcaggg
     1021 ggcggacagc gtggagccca tgttccggca cctgaagaac acgtatgcgg gcctgcagct
     1081 ggtggtggtc atcctgcccg gcaagacgcc cgtgtacgcc gaggtcaagc gcgtgggaga
     1141 cacggtgctg gggatggcca cgcagtgcgt gcagatgaag aacgtgcaga ggaccacgcc
     1201 acagaccctg tccaacctct gcctgaagat caacgtcaag ctgggaggcg tgaacaacat
     1261 cctgctgccc cagggcaggc cgccggtgtt ccagcagccc gtcatctttc tgggagcaga
     1321 cgtcactcac ccccccgccg gggatgggaa gaagccctcc attgccgccg tggtgggcag
     1381 catggacgcc caccccaatc gctactgcgc caccgtgcgc gtgcagcagc accggcagga
     1441 gatcatacaa gacctggccg ccatggtccg cgagctcctc atccagttct acaagtccac
     1501 gcgcttcaag cccacccgca tcatcttcta ccgcgacggt gtctctgaag gccagttcca
     1561 gcaggttctc caccacgagt tgctggccat ccgtgaggcc tgtatcaagc tagaaaaaga
     1621 ctaccagccc gggatcacct tcatcgtggt gcagaagagg caccacaccc ggctcttctg
     1681 cactgacaag aacgagcggg ttgggaaaag tggaaacatt ccagcaggca cgactgtgga
     1741 cacgaaaatc acccacccca ccgagttcga cttctacctg tgtagtcacg ctggcatcca
     1801 ggggacaagc aggccttcgc actatcacgt cctctgggac gacaatcgtt tctcctctga
     1861 tgagctgcag atcctaacct accagctgtg tcacacctac gtgcgctgca cacgctccgt
     1921 gtccatccca gcgccagcat actacgctca cctggtggcc ttccgggcca ggtaccacct
     1981 ggtggataag gaacatgaca gtgctgaagg aagccatacc tctgggcaga gtaacgggcg
     2041 agaccaccaa gcactggcca aggcggtcca ggttcaccaa gacactctgc gcaccatgta
     2101 ctttgcttga cggaattcag tggatccact agtaacggcc gccagtgtgc tggaattaat
     2161 tcgctgtctg cgagggccag ctgttggggt gagtactccc tctcaaaagc gggcatgact
     2221 tctgcgctaa gattgtcagt ttccaaaaac gaggaggatt tgatattcac ctggcccgcg
     2281 gtgatgcctt tgagggtggc cgcgtccatc tggtcagaaa agacaatctt tttgttgtca
     2341 agcttgaggt gtggcaggct tgagatctgg ccatacactt gagtgacaat gacatccact
     2401 ttgcctttct ctccacaggt gtccactccc aggtccaact gcaggtcgag catgcatcta
     2461 gggcggccaa ttccgcccct cctccccccc cccctcctcc cctccccccc cctaacgtta
     2521 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     2581 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     2641 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     2701 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     2761 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     2821 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     2881 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     2941 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     3001 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     3061 gataagcttg ccacaacccg ggataattcc tgcagccaat atgggatcgg ccattgaaca
     3121 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3181 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3241 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3301 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3361 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3421 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3481 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3541 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3601 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgatgatct
     3661 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3721 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3781 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3841 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3901 ctgaggggat caattctcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     3961 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4021 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4081 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4141 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgag
     4201 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4261 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4321 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4381 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4441 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4501 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4561 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4621 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4681 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4741 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4801 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4861 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4921 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4981 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5041 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5101 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5161 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5221 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5281 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5341 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5401 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5461 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5521 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5581 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5641 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5701 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5761 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5821 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5881 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5941 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6001 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6061 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6121 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6181 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6241 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6301 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6361 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6421 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6481 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6541 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     6601 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     6661 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     6721 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6781 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6841 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6901 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6961 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     7021 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     7081 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     7141 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     7201 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     7261 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     7321 tcactatagg gagacccaag cttggtaccg agctcggatc gatatcgccg gccgccatgg
     7381 actacaagga cgacgatgac aagtaccctt atgacgtgcc cgattacgct agcggccgca
     7441 tgtactcggg agccggcccc gcacttgcac ctcctgcgcc gccgcccccc atccaaggat
     7501 atgccttcaa gcctccacct agacccgact ttgggacctc cgggagaaca atcaaattac
     7561 aggccaattt cttcgaaatg gacatcccca aaattgacat ctatcattat gaattggata
     7621 tcaagccaga gaagtgcccg aggagagtta acagggaaat cgtggaacac atggtccagc
     7681 actttaaaac acagatcttt ggggatcgga agcccgtgtt tgacggcagg aagaatctat
     7741 acacagccat gccccttccg attgggaggg acaaggtgga gctggaggtc acgctgccag
     7801 gagaaggcaa ggatcgcatc ttcaaggtgt ccatcaagtg ggtgtcctgc gtgagcttgc
     7861 aggcgttaca cgatgcactt tcagggcggc tgcccagcgt cccttttga
//
