LOCUS       sequence-6799-ex        7893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7893)
  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 7893)
  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
FEATURES             Location/Qualifiers
     source          1..7893
                     /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      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             930..953
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             954..980
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2919..2928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3649..3878
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3861..3880
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3936..4510,4511..4513,4514..4522)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4103..4120)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4330..4349
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4545..5348
                     /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(4608..4627)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5218..5237
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5380..5397)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5386..5610
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5657..5676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5736..5752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5736..5752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5749..5771)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5760..5776
                     /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(5784..5790,5791..5808,5809..5814))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5829..5850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5967..5984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6138..6726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6218..6237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6897..7688,7689..7757))
                     /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     7520..7539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7758..7862)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gccgccatgg actacaagga cgacgatgac aagtaccctt
      961 atgacgtgcc cgattacgct agcggccgca tgggtgttct ctctgccatt cccgcacttg
     1021 cacctcctgc gccgccgccc cccatccaag gatatgcctt caagcctcca cctagacccg
     1081 actttgggac ctccgggaga acaatcaaat tacaggccaa tttcttcgaa atggacatcc
     1141 ccaaaattga catctatcat tatgaattgg atatcaagcc agagaagtgc ccgaggagag
     1201 ttaacaggga aatcgtggaa cacatggtcc agcactttaa aacacagatc tttggggatc
     1261 ggaagcccgt gtttgacggc aggaagaatc tatacacagc catgcccctt ccgattggga
     1321 gggacaaggt ggagctggag gtcacgctgc caggagaagg caaggatcgc atcttcaagg
     1381 tgtccatcaa gtgggtgtcc tgcgtgagct tgcaggcgtt acacgatgca ctttcagggc
     1441 ggctgcccag cgtccctttt gagacgatcc aggccctgga cgtggtcatg aggcacttgc
     1501 catccatgag gtacaccccc gtgggccgct ccttcttcac cgcgtccgaa ggctgctcta
     1561 accctcttgg cgggggccga gaagtgtggt ttggcttcca tcagtccgtc cggccttctc
     1621 tctggaaaat gatgctgaat attgatgtgt cagcaacagc gttttacaag gcacagccag
     1681 taatcgagtt tgtttgtgaa gttttggatt ttaaaagtat tgaagaacaa caaaaacctc
     1741 tgacagattc ccaaagggta aagtttacca aagaaattaa aggtctaaag gtggagataa
     1801 cgcactgtgg gcagatgaag aggaagtacc gtgtctgcaa tgtgacccgg cggcccgcca
     1861 gtcaccaaac attcccgctg cagcaggaga gcgggcagac ggtggagtgc acggtggccc
     1921 agtatttcaa ggacaggcac aagttggttc tgcgctaccc ccacctccca tgtttacaag
     1981 tcggacagga gcagaaacac acctaccttc ccctggaggt ctgtaacatt gtggcaggac
     2041 aaagatgtat taaaaaatta acggacaatc agacctcaac catgatcaga gcaactgcta
     2101 ggtcggcgcc cgatcggcaa gaagagatta gcaaattgat gcgaagtgca agtttcaaca
     2161 cagatccata cgtccgtgaa tttggaatca tggtcaaaga tgagatgaca gacgtgactg
     2221 ggcgggtgct gcagccgccc tccatcctct acgggggcag gaataaagct attgcgaccc
     2281 ctgtccaggg cgtctgggac atgcggaaca agcagttcca cacgggcatc gagatcaagg
     2341 tgtgggccat tgcgtgcttc gccccccagc gccagtgcac ggaagtccat ctgaagtcct
     2401 tcacagagca gctcagaaag atctcgagag acgctggcat gcccatccag ggccagccgt
     2461 gcttctgcaa atacgcgcag ggggcggaca gcgtggagcc catgttccgg cacctgaaga
     2521 acacgtatgc gggcctgcag ctggtggtgg tcatcctgcc cggcaagacg cccgtgtacg
     2581 ccgaggtcaa gcgcgtggga gacacggtgc tggggatggc cacgcagtgc gtgcagatga
     2641 agaacgtgca gaggaccacg ccacagaccc tgtccaacct ttgcctgaag atcaacgtca
     2701 agctgggagg cgtgaacaac atcctgctgc cccagggcag gccgccggtg ttccagcagc
     2761 ccgtcatctt tctgggagca gacgtcactc acccccccgc cggggatggg aagaagccct
     2821 ccattgccgc cgtggtgggc agcatggacg cccaccccaa tcgctactgc gccaccgtgc
     2881 gcgtgcagca gcaccggcag gagatcatac aagacctggc cgccatggtc cgcgagctcc
     2941 tcatccagtt ctacaagtcc acgcgcttca agcccacccg catcatcttc taccgcgacg
     3001 gtgtctctga aggccagttc cagcaggttc tccaccacga gttgctggcc atccgtgagg
     3061 cctgtatcaa gctagaaaaa gactaccagc ccgggatcac cttcatcgtg gtgcagaaga
     3121 ggcaccacac ccggctcttc tgcactgaca agaacgagcg ggttgggaaa agtggaaaca
     3181 ttccagcagg cacgactgtg gacacgaaaa tcacccaccc caccgagttc gacttctacc
     3241 tgtgtagtca cgctggcatc caggggacaa gcaggccttc gcactatcac gtcctctggg
     3301 acgacaatcg tttctcctct gatgagctgc agatcctaac ctaccagctg tgtcacacct
     3361 acgtgcgctg cacacgctcc gtgtccatcc cagcgccagc atactacgct cacctggtgg
     3421 ccttccgggc caggtaccac ctggtggata aggaacatga cagtgctgaa ggaagccata
     3481 cctctgggca gagtaacggg cgagaccacc aagcactggc caaggcggtc caggttcacc
     3541 aagacactct gcgcaccatg tactttgctt gagaattcag tggatccact agtaacggcc
     3601 gccagtgtgc tggaattaat tcgctgtctg cgagggccag ctgttggggt gagtactccc
     3661 tctcaaaagc gggcatgact tctgcgctaa gattgtcagt ttccaaaaac gaggaggatt
     3721 tgatattcac ctggcccgcg gtgatgcctt tgagggtggc cgcgtccatc tggtcagaaa
     3781 agacaatctt tttgttgtca agcttgaggt gtggcaggct tgagatctgg ccatacactt
     3841 gagtgacaat gacatccact ttgcctttct ctccacaggt gtccactccc aggtccaact
     3901 gcaggtcgag catgcatcta gggcggccaa ttccgcccct ctccctcccc cccccctaac
     3961 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gtgattttcc
     4021 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     4081 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     4141 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     4201 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     4261 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     4321 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     4381 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     4441 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     4501 cgatgataag cttgccacaa cccgggataa ttcctgcagc caatatggga tcggccattg
     4561 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4621 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4681 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4741 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4801 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4861 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4921 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4981 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5041 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgatg
     5101 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5161 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5221 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5281 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5341 tcttctgagg ggatcaattc tctagagctc gctgatcagc ctcgactgtg ccttctagtt
     5401 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5461 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5521 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     5581 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     5641 cgagtgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5701 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5761 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5821 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5881 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5941 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6001 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6061 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6121 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6181 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6241 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6301 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     6361 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6421 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6481 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6541 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6601 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6661 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6721 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6781 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6841 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6901 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6961 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7021 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7081 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7141 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7201 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7261 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7321 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7381 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7441 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7501 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7561 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7621 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7681 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7741 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7801 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7861 cgcacatttc cccgaaaagt gccacctgac gtc
//
