LOCUS       sequence-223065-        8900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector to produce NHA-tagged human Ago2 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8900)
  AUTHORS   Pillai RS, Artus CG, Filipowicz W
  TITLE     Tethering of human Ago proteins to mRNA mimics the miRNA-mediated 
            repression of protein synthesis.
  JOURNAL   RNA. 2004 Oct . 10(10):1518-25.
  PUBMED    15337849
REFERENCE   2  (bases 1 to 8900)
  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: "RNA. 2004 
            Oct . 10(10):1518-25."
COMMENT     Sequence Label: pCIneo-NHA-Ago2
FEATURES             Location/Qualifiers
     source          1..8900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        800..1179
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1180..1391
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1342..1362
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1388..1412
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1552..1684
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1667..1686
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1729..1747
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1751..1760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1757..1822
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1859..1885
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1865..1891
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     regulatory      3851..3860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5266..5387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5277..5296)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5331..5350
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5573..6028
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5660..5679)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5870..5891
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6145..6502
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6145..6336
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6168..6188)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6353..6488
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6553..7347
                     /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(6607..6626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7217..7236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7411..7459
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(7500..7519)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7619..7641
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7679..7697)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7765..7869
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7870..7938,7939..8730)
                     /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(8088..8107)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
      661 cttcaatatt ggccattagc catattattc attggttata tagcataaat caatattggc
      721 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
      781 ccaatatgac cgccatgttg gcattgatta ttgactagtt attaatagta atcaattacg
      841 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      901 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      961 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     1021 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
     1081 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
     1141 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     1201 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     1261 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     1321 tgcgatcgcc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1381 taagcagagc tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc
     1441 acagttaaat tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca
     1501 gtgactctct taaggtagcc ttgcagaagt tggtcgtgag gcactgggca ggtaagtatc
     1561 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
     1621 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
     1681 acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta atacgactca
     1741 ctataggcta gccaccatgg acgcacaaac acgacgacgt gagcgtcgcg ctgagaaaca
     1801 agctcaatgg aaagctgcaa acccaccgct cgaggctgcc caagcttaca ccatggccta
     1861 cccatatgat gttccagatt acgcttcact cgacgcggat cccgtcgaat tcaccatggg
     1921 agggcaccca ttccagtggt gtaacgcact tgcacctcct gcgccgccgc cccccatcca
     1981 aggatatgcc ttcaagcctc cacctagacc cgactttggg acctccggga gaacaatcaa
     2041 attacaggcc aatttcttcg aaatggacat ccccaaaatt gacatctatc attatgaatt
     2101 ggatatcaag ccagagaagt gcccgaggag agttaacagg gaaatcgtgg aacacatggt
     2161 ccagcacttt aaaacacaga tctttgggga tcggaagccc gtgtttgacg gcaggaagaa
     2221 tctatacaca gccatgcccc ttccgattgg gagggacaag gtggagctgg aggtcacgct
     2281 gccaggagaa ggcaaggatc gcatcttcaa ggtgtccatc aagtgggtgt cctgcgtgag
     2341 cttgcaggcg ttacacgatg cactttcagg gcggctgccc agcgtccctt ttgagacgat
     2401 ccaggccctg gacgtggtca tgaggcactt gccatccatg aggtacaccc ccgtgggccg
     2461 ctccttcttc accgcgtccg aaggctgctc taaccctctt ggcgggggcc gagaagtgtg
     2521 gtttggcttc catcagtccg tccggccttc tctctggaaa atgatgctga atattgatgt
     2581 gtcagcaaca gcgttttaca aggcacagcc agtaatcgag tttgtttgtg aagttttgga
     2641 ttttaaaagt attgaagaac aacaaaaacc tctgacagat tcccaaaggg taaagtttac
     2701 caaagaaatt aaaggtctaa aggtggagat aacgcactgt gggcagatga agaggaagta
     2761 ccgtgtctgc aatgtgaccc ggcggcccgc cagtcaccaa acattcccgc tgcagcagga
     2821 gagcgggcag acggtggagt gcacggtggc ccagtatttc aaggacaggc acaagttggt
     2881 tctgcgctac ccccacctcc catgtttaca agtcggacag gagcagaaac acacctacct
     2941 tcccctggag gtctgtaaca ttgtggcagg acaaagatgt attaaaaaat taacggacaa
     3001 tcagacctca accatgatca gagcaactgc taggtcggcg cccgatcggc aagaagagat
     3061 tagcaaattg atgcgaagtg caagtttcaa cacagatcca tacgtccgtg aatttggaat
     3121 catggtcaaa gatgagatga cagacgtgac tgggcgggtg ctgcagccgc cctccatcct
     3181 ctacgggggc aggaataaag ctattgcgac ccctgtccag ggcgtctggg acatgcggaa
     3241 caagcagttc cacacgggca tcgagatcaa ggtgtgggcc attgcgtgct tcgcccccca
     3301 gcgccagtgc acggaagtcc atctgaagtc cttcacagag cagctcagaa agatctcgag
     3361 agacgctggc atgcccatcc agggccagcc gtgcttctgc aaatacgcgc agggggcgga
     3421 cagcgtggag cccatgttcc ggcacctgaa gaacacgtat gcgggcctgc agctggtggt
     3481 ggtcatcctg cccggcaaga cgcccgtgta cgccgaggtc aagcgcgtgg gagacacggt
     3541 gctggggatg gccacgcagt gcgtgcagat gaagaacgtg cagaggacca cgccacagac
     3601 cctgtccaac ctctgcctga agatcaacgt caagctggga ggcgtgaaca acatcctgct
     3661 gccccagggc aggccgccgg tgttccagca gcccgtcatc tttctgggag cagacgtcac
     3721 tcaccccccc gccggggatg ggaagaagcc ctccattgcc gccgtggtgg gcagcatgga
     3781 cgcccacccc aatcgctact gcgccaccgt gcgcgtgcag cagcaccggc aggagatcat
     3841 acaagacctg gccgccatgg tccgcgagct cctcatccag ttctacaagt ccacgcgctt
     3901 caagcccacc cgcatcatct tctaccgcga cggtgtctct gaaggccagt tccagcaggt
     3961 tctccaccac gagttgctgg ccatccgtga ggcctgtatc aagctagaaa aagactacca
     4021 gcccgggatc accttcatcg tggtgcagaa gaggcaccac acccggctct tctgcactga
     4081 caagaacgag cgggttggga aaagtggaaa cattccagca ggcacgactg tggacacgaa
     4141 aatcacccac cccaccgagt tcgacttcta cctgtgtagt cacgctggca tccaggggac
     4201 aagcaggcct tcgcactatc acgtcctctg ggacgacaat cgtttctcct ctgatgagct
     4261 gcagatccta acctaccagc tgtgtcacac ctacgtgcgc tgcacacgct ccgtgtccat
     4321 cccagcgcca gcatactacg ctcacctggt ggccttccgg gccaggtacc acctggtgga
     4381 taaggaacat gacagtgctg aaggaagcca tacctctggg cagagtaacg ggcgagacca
     4441 ccaagcactg gccaaggcgg tccaggttca ccaagacact ctgcgcacca tgtactttgc
     4501 ttgacatgtt ttagtgttta gcgattgtgt accgagtggg attcacgaga ccagctacac
     4561 tcagaccaac agatggccag cccttccgtg acagccagca tcgaacatga gacgtcattg
     4621 attttattag attctccgtt ttccagaatg ccttccgtcc cagatttcaa acttggattt
     4681 tgaactgcag acctgtatga gaacccaatg tcataggaaa tatggtttgc taaaatctat
     4741 aagctgctta ttaaaacaga gtcccgtgtg tcctaaaaat ctcctaaaac cagtctatga
     4801 actcagggct ttaaaacatt tttaatttat ttggtcattc aatttacttg tttttaatac
     4861 atgattctct atgaaattga tgggctcaaa ctagctgtga atcttctgag agtgaaagca
     4921 acacaaaaca caagtgtggt tttaaagcct tgaacattct gatgttgtca ctaaagttga
     4981 tttccaggcg atgctcgtgt gcccctggcg tggctcaccc aagttcctcg actgagggcc
     5041 ggtggccatg cagaggcgtc cgcaggtgcc gtgttctgcc agcaccgccc ttcacccggc
     5101 ctgaactaag gagcagtggc agaaggtggg ccccgtgtgt ttacagcatt tccaggtcca
     5161 gagaggttgg cagacaagtg ccattttaat aaaaaatgta tttaaaaaaa aaaaaaaggg
     5221 cggccgcttc cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg
     5281 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     5341 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     5401 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     5461 acctctacaa atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga
     5521 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     5581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5641 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5701 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5761 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5881 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5941 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6001 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     6061 gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga
     6121 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     6181 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6241 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6301 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6361 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6421 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6481 tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag
     6541 gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6601 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6661 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6721 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6781 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6841 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6901 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6961 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     7021 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     7081 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     7141 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     7201 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     7261 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     7321 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     7381 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     7441 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt
     7501 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     7561 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     7621 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     7681 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     7741 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7801 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7861 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7921 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7981 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     8041 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     8101 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     8161 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     8221 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     8281 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     8341 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     8401 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     8461 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     8521 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     8581 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     8641 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     8701 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     8761 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8821 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8881 gaaaagatca aaggatcttc
//
