LOCUS       sequence-288733-        8185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAGO2-F470V.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8185)
  AUTHORS   Eulalio A, Huntzinger E, Izaurralde E
  TITLE     GW182 interaction with Argonaute is essential for miRNA-mediated 
            translational repression and mRNA decay.
  JOURNAL   Nat Struct Mol Biol. 2008 Apr . 15(4):346-53.
  PUBMED    18345015
REFERENCE   2  (bases 1 to 8185)
  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: "Nat Struct 
            Mol Biol. 2008 Apr . 15(4):346-53."
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsAGO2-F470V_F
FEATURES             Location/Qualifiers
     source          1..8185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1089..1098
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1095..1160
                     /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             1197..1223
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1203..1229
                     /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      3189..3198
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3889..4010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3900..3919)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3954..3973
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4196..4651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4283..4302)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4493..4514
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4768..5125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4768..4959
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4791..4811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4976..5111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5176..5970
                     /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(5230..5249)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5840..5859
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6034..6082
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6123..6142)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6242..6264
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6302..6320)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6388..6492
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6493..6561,6562..7353)
                     /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(6711..6730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7524..8112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8013..8032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag
     1141 ctcaatggaa agctgcaaac ccaccgctcg aggctgccca agcttacacc atggcctacc
     1201 catatgatgt tccagattac gcttcactcg acgcggatcc cgtcgaattc accatgggag
     1261 ggcacccatt ccagtggtgt aacgcacttg cacctcctgc gccgccgccc cccatccaag
     1321 gatatgcctt caagcctcca cctagacccg actttgggac ctccgggaga acaatcaaat
     1381 tacaggccaa tttcttcgaa atggacatcc ccaaaattga catctatcat tatgaattgg
     1441 atatcaagcc agagaagtgc ccgaggagag ttaacaggga aatcgtggaa cacatggtcc
     1501 agcactttaa aacacagatc tttggggatc ggaagcccgt gtttgacggc aggaagaatc
     1561 tatacacagc catgcccctt ccgattggga gggacaaggt ggagctggag gtcacgctgc
     1621 caggagaagg caaggatcgc atcttcaagg tgtccatcaa gtgggtgtcc tgcgtgagct
     1681 tgcaggcgtt acacgatgca ctttcagggc ggctgcccag cgtccctttt gagacgatcc
     1741 aggccctgga cgtggtcatg aggcacttgc catccatgag gtacaccccc gtgggccgct
     1801 ccttcttcac cgcgtccgaa ggctgctcta accctcttgg cgggggccga gaagtgtggt
     1861 ttggcttcca tcagtccgtc cggccttctc tctggaaaat gatgctgaat attgatgtgt
     1921 cagcaacagc gttttacaag gcacagccag taatcgagtt tgtttgtgaa gttttggatt
     1981 ttaaaagtat tgaagaacaa caaaaacctc tgacagattc ccaaagggta aagtttacca
     2041 aagaaattaa aggtctaaag gtggagataa cgcactgtgg gcagatgaag aggaagtacc
     2101 gtgtctgcaa tgtgacccgg cggcccgcca gtcaccaaac attcccgctg cagcaggaga
     2161 gcgggcagac ggtggagtgc acggtggccc agtatttcaa ggacaggcac aagttggttc
     2221 tgcgctaccc ccacctccca tgtttacaag tcggacagga gcagaaacac acctaccttc
     2281 ccctggaggt ctgtaacatt gtggcaggac aaagatgtat taaaaaatta acggacaatc
     2341 agacctcaac catgatcaga gcgactgcta ggtcggcgcc cgatcggcaa gaagagatta
     2401 gcaaattgat gcgaagtgca agtttcaaca cagatccata cgtccgtgaa tttggaatca
     2461 tggtcaaaga tgagatgaca gacgtgactg ggcgggtgct gcagccgccc tccatcctct
     2521 acgggggcag gaataaagct attgcgaccc ctgtccaggg cgtctgggac atgcggaaca
     2581 agcagttcca cacgggcatc gagatcaagg tgtgggccat tgcgtgcttc gccccccagc
     2641 gccagtgcac ggaagtccat ctgaagtccg tcacagagca gctcagaaag atctcgagag
     2701 acgccggcat gcccatccag ggccagccgt gcttctgcaa atacgcgcag ggggcggaca
     2761 gcgtggagcc catgttccgg cacctgaaga acacgtatgc gggcctgcag ctggtggtgg
     2821 tcatcctgcc cggcaagacg cccgtgtacg ccgaggtcaa gcgcgtggga gacacggtgc
     2881 tggggatggc cacgcagtgc gtgcagatga agaacgtgca gaggaccacg ccacagaccc
     2941 tgtccaacct ctgcctgaag atcaacgtca agctgggagg cgtgaacaac atcctgctgc
     3001 cccagggcag gccgccggtg ttccagcagc ccgtcatctt tctgggagca gacgtcactc
     3061 acccccccgc cggggatggg aagaagccct ccattgccgc cgtggtgggc agcatggacg
     3121 cccaccccaa tcgctactgc gccaccgtgc gcgtgcagca gcaccggcag gagatcatac
     3181 aagacctggc cgccatggtc cgcgagctcc tcatccagtt ctacaagtcc acgcgcttca
     3241 agcccacccg catcatcttc taccgcgacg gtgtctctga aggccagttc cagcaggttc
     3301 tccaccacga gttgctggcc atccgtgagg cctgtatcaa gctagaaaaa gactaccagc
     3361 ccgggatcac cttcatcgtg gtgcagaaga ggcaccacac ccggctcttc tgcactgaca
     3421 agaacgagcg ggttgggaaa agtggaaaca ttccagcagg cacgactgtg gacacgaaaa
     3481 tcacccaccc caccgagttc gacttctacc tgtgtagtca cgctggcatc caggggacaa
     3541 gcaggccttc gcactatcac gtcctctggg acgacaatcg tttctcctct gatgagctgc
     3601 agatcctaac ctaccagctg tgtcacacct acgtgcgctg cacacgctcc gtgtccatcc
     3661 cagcgccagc atactacgct cacctggtgg ccttccgggc caggtaccac ctggtggata
     3721 aggaacatga cagtgctgaa ggaagccata cctctgggca gagtaacggg cgagaccacc
     3781 aagcactggc caaggcggtc caggttcacc aagacactct gcgcaccatg tactttgctt
     3841 gagcggccgc ttccctttag tgagggttaa tgcttcgagc agacatgata agatacattg
     3901 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3961 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4021 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     4081 aaaacctcta caaatgtggt aaaatccgat aaggatcgat ccgggctggc gtaatagcga
     4141 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     4201 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4261 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4321 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4381 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4441 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4501 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4561 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4621 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     4681 tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc
     4741 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     4801 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4861 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4921 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4981 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5041 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5101 ttttggaggc ctaggctttt gcaaaaagct tgattcttct gacacaacag tctcgaactt
     5161 aaggctagag ccaccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5221 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5281 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5341 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5401 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5461 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5521 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5581 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5641 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5701 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5761 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5821 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5881 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5941 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6001 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     6061 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgcgtatg gtgcactctc
     6121 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6181 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6241 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     6301 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6361 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6421 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6481 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6541 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6601 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6661 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6721 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6781 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6841 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6901 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6961 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7021 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7081 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7141 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7201 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7261 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7321 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7381 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7441 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7501 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7561 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7621 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7681 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7741 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7801 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7861 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7921 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7981 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8041 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8101 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8161 tttgctcaca tggctcgaca gatct
//
