LOCUS       sequence-6800-ex        7890 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 7890)
  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 7890)
  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..7890
                     /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      2916..2925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3646..3875
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3858..3877
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3933..4507,4508..4510,4511..4519)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4100..4117)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4327..4346
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4542..5345
                     /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(4605..4624)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5215..5234
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5377..5394)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5383..5607
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5654..5673
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5733..5749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5733..5749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5746..5768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5757..5773
                     /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(5781..5787,5788..5805,5806..5811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5826..5847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5964..5981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6135..6723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6215..6234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6894..7685,7686..7754))
                     /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     7517..7536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7755..7859)
                     /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 tgtactcggg agccggcccc gcacttgcac
     1021 ctcctgcgcc gccgcccccc atccaaggat atgccttcaa gcctccacct agacccgact
     1081 ttgggacctc cgggagaaca atcaaattac aggccaattt cttcgaaatg gacatcccca
     1141 aaattgacat ctatcattat gaattggata tcaagccaga gaagtgcccg aggagagtta
     1201 acagggaaat cgtggaacac atggtccagc actttaaaac acagatcttt ggggatcgga
     1261 agcccgtgtt tgacggcagg aagaatctat acacagccat gccccttccg attgggaggg
     1321 acaaggtgga gctggaggtc acgctgccag gagaaggcaa ggatcgcatc ttcaaggtgt
     1381 ccatcaagtg ggtgtcctgc gtgagcttgc aggcgttaca cgatgcactt tcagggcggc
     1441 tgcccagcgt cccttttgag acgatccagg ccctggacgt ggtcatgagg cacttgccat
     1501 ccatgaggta cacccccgtg ggccgctcct tcttcaccgc gtccgaaggc tgctctaacc
     1561 ctcttggcgg gggccgagaa gtgtggtttg gcttccatca gtccgtccgg ccttctctct
     1621 ggaaaatgat gctgaatatt gatgtgtcag caacagcgtt ttacaaggca cagccagtaa
     1681 tcgagtttgt ttgtgaagtt ttggatttta aaagtattga agaacaacaa aaacctctga
     1741 cagattccca aagggtaaag tttaccaaag aaattaaagg tctaaaggtg gagataacgc
     1801 actgtgggca gatgaagagg aagtaccgtg tctgcaatgt gacccggcgg cccgccagtc
     1861 accaaacatt cccgctgcag caggagagcg ggcagacggt ggagtgcacg gtggcccagt
     1921 atttcaagga caggcacaag ttggttctgc gctaccccca cctcccatgt ttacaagtcg
     1981 gacaggagca gaaacacacc taccttcccc tggaggtctg taacattgtg gcaggacaaa
     2041 gatgtattaa aaaattaacg gacaatcaga cctcaaccat gatcagagcg actgctaggt
     2101 cggcgcccga tcggcaagaa gagattagca aattgatgcg aagtgcaagt ttcaacacag
     2161 atccatacgt ccgtgaattt ggaatcatgg tcaaagatga gatgacagac gtgactgggc
     2221 gggtgctgca gccgccctcc atcctctacg ggggcaggaa taaagctatt gcgacccctg
     2281 tccagggcgt ctgggacatg cggaacaagc agttccacac gggcatcgag atcaaggtgt
     2341 gggccattgc gtgcttcgcc ccccagcgcc agtgcacgga agtccatctg aagtccttca
     2401 cagagcagct cagaaagatc tcgagagacg ccggcatgcc catccagggc cagccgtgct
     2461 tctgcaaata cgcgcagggg gcggacagcg tggagcccat gttccggcac ctgaagaaca
     2521 cgtatgcggg cctgcagctg gtggtggtca tcctgcccgg caagacgccc gtgtacgccg
     2581 aggtcaagcg cgtgggagac acggtgctgg ggatggccac gcagtgcgtg cagatgaaga
     2641 acgtgcagag gaccacgcca cagaccctgt ccaacctctg cctgaagatc aacgtcaagc
     2701 tgggaggcgt gaacaacatc ctgctgcccc agggcaggcc gccggtgttc cagcagcccg
     2761 tcatctttct gggagcagac gtcactcacc cccccgccgg ggatgggaag aagccctcca
     2821 ttgccgccgt ggtgggcagc atggacgccc accccaatcg ctactgcgcc accgtgcgcg
     2881 tgcagcagca ccggcaggag atcatacaag acctggccgc catggtccgc gagctcctca
     2941 tccagttcta caagtccacg cgcttcaagc ccacccgcat catcttctac cgcgacggtg
     3001 tctctgaagg ccagttccag caggttctcc accacgagtt gctggccatc cgtgaggcct
     3061 gtatcaagct agaaaaagac taccagcccg ggatcacctt catcgtggtg cagaagaggc
     3121 accacacccg gctcttctgc actgacaaga acgagcgggt tgggaaaagt ggaaacattc
     3181 cagcaggcac gactgtggac acgaaaatca cccaccccac cgagttcgac ttctacctgt
     3241 gtagtcacgc tggcatccag gggacaagca ggccttcgca ctatcacgtc ctctgggacg
     3301 acaatcgttt ctcctctgat gagctgcaga tcctaaccta ccagctgtgt cacacctacg
     3361 tgcgctgcac acgctccgtg tccatcccag cgccagcata ctacgctcac ctggtggcct
     3421 tccgggccag gtaccacctg gtggataagg aacatgacag tgctgaagga agccatacct
     3481 ctgggcagag taacgggcga gaccaccaag cactggccaa ggcggtccag gttcaccaag
     3541 acactctgcg caccatgtac tttgcttgag aattcagtgg atccactagt aacggccgcc
     3601 agtgtgctgg aattaattcg ctgtctgcga gggccagctg ttggggtgag tactccctct
     3661 caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga
     3721 tattcacctg gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga
     3781 caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag
     3841 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca
     3901 ggtcgagcat gcatctaggg cggccaattc cgcccctctc cctccccccc ccctaacgtt
     3961 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtg attttccacc
     4021 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     4081 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     4141 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     4201 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     4261 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     4321 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     4381 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     4441 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     4501 tgataagctt gccacaaccc gggataattc ctgcagccaa tatgggatcg gccattgaac
     4561 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4621 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4681 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4741 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4801 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4861 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4921 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4981 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5041 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgatgatc
     5101 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5161 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5221 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5281 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5341 tctgagggga tcaattctct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     5401 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5461 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5521 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     5581 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     5641 gtgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5701 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5761 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5821 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5881 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5941 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6001 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6061 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6121 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6181 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6241 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6301 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6361 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6421 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6481 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6541 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6601 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6661 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6721 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6781 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6841 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6901 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6961 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7021 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7081 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7141 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7201 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7261 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7321 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7381 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7441 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7501 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7561 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7621 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7681 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7741 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7801 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7861 acatttcccc gaaaagtgcc acctgacgtc
//
