LOCUS       sequence-223066-        8822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector to produce HA-tagged human Ago2 protein without 
            N-peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8822)
  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 8822)
  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-HA-Ago2
FEATURES             Location/Qualifiers
     source          1..8822
                     /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"
     CDS             1781..1807
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1787..1813
                     /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      3773..3782
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5188..5309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5199..5218)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5253..5272
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5495..5950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5582..5601)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5792..5813
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6067..6424
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6067..6258
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6090..6110)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6275..6410
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6475..7269
                     /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(6529..6548)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7139..7158
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7333..7381
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(7422..7441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7541..7563
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7601..7619)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7687..7791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7792..7860,7861..8652)
                     /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(8010..8029)
                     /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 gtcgaggctg cccaagctta caccatggcc tacccatatg atgttccaga
     1801 ttacgcttca ctcgacgcgg atcccgtcga attcaccatg ggagggcacc cattccagtg
     1861 gtgtaacgca cttgcacctc ctgcgccgcc gccccccatc caaggatatg ccttcaagcc
     1921 tccacctaga cccgactttg ggacctccgg gagaacaatc aaattacagg ccaatttctt
     1981 cgaaatggac atccccaaaa ttgacatcta tcattatgaa ttggatatca agccagagaa
     2041 gtgcccgagg agagttaaca gggaaatcgt ggaacacatg gtccagcact ttaaaacaca
     2101 gatctttggg gatcggaagc ccgtgtttga cggcaggaag aatctataca cagccatgcc
     2161 ccttccgatt gggagggaca aggtggagct ggaggtcacg ctgccaggag aaggcaagga
     2221 tcgcatcttc aaggtgtcca tcaagtgggt gtcctgcgtg agcttgcagg cgttacacga
     2281 tgcactttca gggcggctgc ccagcgtccc ttttgagacg atccaggccc tggacgtggt
     2341 catgaggcac ttgccatcca tgaggtacac ccccgtgggc cgctccttct tcaccgcgtc
     2401 cgaaggctgc tctaaccctc ttggcggggg ccgagaagtg tggtttggct tccatcagtc
     2461 cgtccggcct tctctctgga aaatgatgct gaatattgat gtgtcagcaa cagcgtttta
     2521 caaggcacag ccagtaatcg agtttgtttg tgaagttttg gattttaaaa gtattgaaga
     2581 acaacaaaaa cctctgacag attcccaaag ggtaaagttt accaaagaaa ttaaaggtct
     2641 aaaggtggag ataacgcact gtgggcagat gaagaggaag taccgtgtct gcaatgtgac
     2701 ccggcggccc gccagtcacc aaacattccc gctgcagcag gagagcgggc agacggtgga
     2761 gtgcacggtg gcccagtatt tcaaggacag gcacaagttg gttctgcgct acccccacct
     2821 cccatgttta caagtcggac aggagcagaa acacacctac cttcccctgg aggtctgtaa
     2881 cattgtggca ggacaaagat gtattaaaaa attaacggac aatcagacct caaccatgat
     2941 cagagcaact gctaggtcgg cgcccgatcg gcaagaagag attagcaaat tgatgcgaag
     3001 tgcaagtttc aacacagatc catacgtccg tgaatttgga atcatggtca aagatgagat
     3061 gacagacgtg actgggcggg tgctgcagcc gccctccatc ctctacgggg gcaggaataa
     3121 agctattgcg acccctgtcc agggcgtctg ggacatgcgg aacaagcagt tccacacggg
     3181 catcgagatc aaggtgtggg ccattgcgtg cttcgccccc cagcgccagt gcacggaagt
     3241 ccatctgaag tccttcacag agcagctcag aaagatctcg agagacgctg gcatgcccat
     3301 ccagggccag ccgtgcttct gcaaatacgc gcagggggcg gacagcgtgg agcccatgtt
     3361 ccggcacctg aagaacacgt atgcgggcct gcagctggtg gtggtcatcc tgcccggcaa
     3421 gacgcccgtg tacgccgagg tcaagcgcgt gggagacacg gtgctgggga tggccacgca
     3481 gtgcgtgcag atgaagaacg tgcagaggac cacgccacag accctgtcca acctctgcct
     3541 gaagatcaac gtcaagctgg gaggcgtgaa caacatcctg ctgccccagg gcaggccgcc
     3601 ggtgttccag cagcccgtca tctttctggg agcagacgtc actcaccccc ccgccgggga
     3661 tgggaagaag ccctccattg ccgccgtggt gggcagcatg gacgcccacc ccaatcgcta
     3721 ctgcgccacc gtgcgcgtgc agcagcaccg gcaggagatc atacaagacc tggccgccat
     3781 ggtccgcgag ctcctcatcc agttctacaa gtccacgcgc ttcaagccca cccgcatcat
     3841 cttctaccgc gacggtgtct ctgaaggcca gttccagcag gttctccacc acgagttgct
     3901 ggccatccgt gaggcctgta tcaagctaga aaaagactac cagcccggga tcaccttcat
     3961 cgtggtgcag aagaggcacc acacccggct cttctgcact gacaagaacg agcgggttgg
     4021 gaaaagtgga aacattccag caggcacgac tgtggacacg aaaatcaccc accccaccga
     4081 gttcgacttc tacctgtgta gtcacgctgg catccagggg acaagcaggc cttcgcacta
     4141 tcacgtcctc tgggacgaca atcgtttctc ctctgatgag ctgcagatcc taacctacca
     4201 gctgtgtcac acctacgtgc gctgcacacg ctccgtgtcc atcccagcgc cagcatacta
     4261 cgctcacctg gtggccttcc gggccaggta ccacctggtg gataaggaac atgacagtgc
     4321 tgaaggaagc catacctctg ggcagagtaa cgggcgagac caccaagcac tggccaaggc
     4381 ggtccaggtt caccaagaca ctctgcgcac catgtacttt gcttgacatg ttttagtgtt
     4441 tagcgattgt gtaccgagtg ggattcacga gaccagctac actcagacca acagatggcc
     4501 agcccttccg tgacagccag catcgaacat gagacgtcat tgattttatt agattctccg
     4561 ttttccagaa tgccttccgt cccagatttc aaacttggat tttgaactgc agacctgtat
     4621 gagaacccaa tgtcatagga aatatggttt gctaaaatct ataagctgct tattaaaaca
     4681 gagtcccgtg tgtcctaaaa atctcctaaa accagtctat gaactcaggg ctttaaaaca
     4741 tttttaattt atttggtcat tcaatttact tgtttttaat acatgattct ctatgaaatt
     4801 gatgggctca aactagctgt gaatcttctg agagtgaaag caacacaaaa cacaagtgtg
     4861 gttttaaagc cttgaacatt ctgatgttgt cactaaagtt gatttccagg cgatgctcgt
     4921 gtgcccctgg cgtggctcac ccaagttcct cgactgaggg ccggtggcca tgcagaggcg
     4981 tccgcaggtg ccgtgttctg ccagcaccgc ccttcacccg gcctgaacta aggagcagtg
     5041 gcagaaggtg ggccccgtgt gtttacagca tttccaggtc cagagaggtt ggcagacaag
     5101 tgccatttta ataaaaaatg tatttaaaaa aaaaaaaaag ggcggccgct tccctttagt
     5161 gagggttaat gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     5221 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5281 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     5341 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     5401 aaatccgata aggatcgatc cgggctggcg taatagcgaa gaggcccgca ccgatcgccc
     5461 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     5521 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5581 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5641 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5701 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5761 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5821 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5881 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5941 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6001 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     6061 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     6121 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6181 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6241 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6301 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6361 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6421 caaaaagctt gattcttctg acacaacagt ctcgaactta aggctagagc caccatgatt
     6481 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     6541 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6601 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     6661 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6721 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6781 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6841 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6901 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6961 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     7021 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     7081 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     7141 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     7201 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     7261 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     7321 cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     7381 gaatcgatag cgataaggat ccgcgtatgg tgcactctca gtacaatctg ctctgatgcc
     7441 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7501 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     7561 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     7621 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     7681 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7741 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7801 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7861 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7921 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7981 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     8041 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8101 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8161 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8221 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8281 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     8341 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8401 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8461 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     8521 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     8581 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8641 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8701 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8821 tc
//
