LOCUS       sequence-476231-        7318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAGO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7318)
  AUTHORS   Lazzaretti D, Tournier I, Izaurralde E
  TITLE     The C-terminal domains of human TNRC6A, TNRC6B, and TNRC6C silence 
            bound transcripts independently of Argonaute proteins.
  JOURNAL   RNA. 2009 Jun . 15(6):1059-66.
  PUBMED    19383768
REFERENCE   2  (bases 1 to 7318)
  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. 2009 
            Jun . 15(6):1059-66."
COMMENT     Sequence Label: pT7-EGFP-C1-HsAGO1_F
FEATURES             Location/Qualifiers
     source          1..7318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(650..671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(911..930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1258..1279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4046..4167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4083..4102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4137..4156
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4174..4629)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4311..4332)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4523..4542
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4656..4760
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4762..5119
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4762..4953
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4785..4805)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4970..5105
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5154..5948
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5154..5948
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5208..5227)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5818..5837
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6136..6155)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6180..6227
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6556..7144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7045..7064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      661 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      721 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      781 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      841 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      901 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      961 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1021 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1081 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1141 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1201 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1261 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1321 gtccggactc agatctcgag ctatggaagc gggaccctcg ggagcagctg cgggcgctta
     1381 cctgcccccc ctgcagcagg tgttccaggc acctcgccgg cctggcattg gcactgtggg
     1441 gaaaccaatc aagctcctgg ccaattactt tgaggtggac atccctaaga tcgacgtgta
     1501 ccactacgag gtggacatca agccggataa gtgtccccgt agagtcaacc gggaagtggt
     1561 ggaatacatg gtccagcatt tcaagcctca gatctttggt gatcgcaagc ctgtgtatga
     1621 tggaaagaag aacatttaca ctgtcacagc actgcccatt ggcaacgaac gggtcgactt
     1681 tgaggtgaca atccctgggg aagggaagga tcgaatcttt aaggtctcca tcaagtggct
     1741 agccattgtg agctggcgaa tgctgcatga ggccctggtc agcggccaga tccctgttcc
     1801 cttggagtct gtgcaagccc tggatgtggc catgaggcac ctggcatcca tgaggtacac
     1861 ccctgtgggc cgctccttct tctcaccgcc tgagggctac taccacccgc tggggggtgg
     1921 gcgcgaggtc tggttcggct ttcaccagtc tgtgcgccct gccatgtgga agatgatgct
     1981 caacattgat gtctcagcca ctgcctttta taaggcacag ccagtgattg agttcatgtg
     2041 tgaggtgctg gacatcagga acatagatga gcagcccaag cccctcacgg actctcagcg
     2101 cgttcgcttc accaaggaga tcaagggcct gaaggtggaa gtcacccact gtggacagat
     2161 gaagaggaag taccgcgtgt gtaatgttac ccgtcgccct gctagccatc agacattccc
     2221 cttacagctg gagagtggac agactgtgga gtgcacagtg gcacagtatt tcaagcagaa
     2281 atataacctt cagctcaagt atccccatct gccctgccta caagttggcc aggaacaaaa
     2341 gcatacctac cttcccctag aggtctgtaa cattgtggct gggcagcgct gtattaaaaa
     2401 gctgaccgac aaccagacct cgaccatgat aaaggccaca gctagatccg ctccagacag
     2461 acaggaggag atcagtcgcc tgatgaagaa tgccagctac aacttagatc cctacatcca
     2521 ggaatttggg atcaaagtga aggatgacat gacggaggtg acagggcgag tgctgccggc
     2581 gcccatcttg cagtacggcg gccggaaccg ggccattgcc acacccaatc agggtgtctg
     2641 ggacatgcgg gggaaacagt tctacaatgg gattgagatc aaagtctggg ccatcgcctg
     2701 cttcgcaccc caaaaacagt gtcgagaaga ggtgctcaag aacttcacag accagctgcg
     2761 gaagatttcc aaggatgcgg ggatgcctat ccagggtcaa ccttgtttct gcaaatatgc
     2821 acagggggca gacagcgtgg agcctatgtt ccggcatctc aagaacacct actcagggct
     2881 gcagctcatt attgtcatcc tgccagggaa gacgccggtg tatgctgagg tgaaacgtgt
     2941 cggagataca ctcttgggaa tggctacgca gtgtgtgcag gtgaagaacg tggtcaagac
     3001 ctcacctcag actctgtcca acctctgcct caagatcaat gtcaaacttg gtggcattaa
     3061 cagcatccta gtcccacacc agcgctctgc cgtttttcaa cagccagtga tattcctggg
     3121 agcagatgtt acacaccccc cagcagggga tgggaaaaaa ccttctatca cagcagtggt
     3181 aggcagtatg gatgcccacc ccagccgata ctgtgctact gtgcgggtac agcgaccacg
     3241 gcaagagatc attgaagact tgtcctacat ggtgcgtgag ctcctcatcc aattctacaa
     3301 gtccacccgt ttcaagccta cccgcatcat cttctaccga gatggggtgc ctgaaggcca
     3361 gctaccccag atactccact atgagctact ggccattcgt gatgcctgca tcaaactgga
     3421 aaaggactac cagcctggga tcacttatat tgtggtgcag aaacgccatc acacccgcct
     3481 tttctgtgct gacaagaatg agcgaattgg gaagagtggt aacatcccag ctgggaccac
     3541 agtggacacc aacatcaccc acccatttga gtttgacttc tatctgtgca gccacgcagg
     3601 catccagggc accagccgac catcccatta ctatgttctt tgggatgaca accgtttcac
     3661 agcagatgag ctccagatcc tgacgtacca gctgtgccac acttacgtac gatgcacacg
     3721 ctctgtctct atcccagcac ctgcctacta tgcccgcctg gtggctttcc gggcacgata
     3781 ccacctggtg gacaaggagc atgacagtgg agaggggagc cacatatcgg ggcagagcaa
     3841 tgggcgggac ccccaggccc tggccaaagc cgtgcaggtt caccaggata ctctgcgcac
     3901 catgtacttc gcttgaggat ccaccggatc tagataactg atcataatca gccataccac
     3961 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4021 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4081 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4141 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     4201 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4261 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4321 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4381 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4441 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4501 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4561 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4621 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4681 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4741 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4801 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4861 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4921 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4981 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5041 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5101 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     5161 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5221 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5281 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5341 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5401 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5461 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5521 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5581 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5641 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5701 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5761 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5821 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5881 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5941 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6001 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6061 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6121 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     6181 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6241 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6301 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6361 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6421 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6481 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6661 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7201 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     7261 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
