LOCUS       sequence-442064-        7423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAGO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7423)
  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 7423)
  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-HsAGO2_F
FEATURES             Location/Qualifiers
     source          1..7423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        313..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     467..487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     561..580
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        561..579
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1330..1395
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      3331..3340
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4159..4280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4196..4215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4250..4269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4287..4742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4424..4445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4636..4655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4769..4873
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4875..5232
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4875..5066
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4898..4918)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5083..5218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5267..6061
                     /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             5267..6061
                     /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(5321..5340)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5931..5950
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6249..6268)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6293..6340
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6669..7257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7158..7177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
       61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      541 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 gcgggcccgg gatccatggg agggcaccca ttccagtggt gtaacgcact tgcacctcct
     1441 gcgccgccgc cccccatcca aggatatgcc ttcaagcctc cacctagacc cgactttggg
     1501 acctccggga gaacaatcaa attacaggcc aatttcttcg aaatggacat ccccaaaatt
     1561 gacatctatc attatgaatt ggatatcaag ccagagaagt gcccgaggag agttaacagg
     1621 gaaatcgtgg aacacatggt ccagcacttt aaaacacaga tctttgggga tcggaagccc
     1681 gtgtttgacg gcaggaagaa tctatacaca gccatgcccc ttccgattgg gagggacaag
     1741 gtggagctgg aggtcacgct gccaggagaa ggcaaggatc gcatcttcaa ggtgtccatc
     1801 aagtgggtgt cctgcgtgag cttgcaggcg ttacacgatg cactttcagg gcggctgccc
     1861 agcgtccctt ttgagacgat ccaggccctg gacgtggtca tgaggcactt gccatccatg
     1921 aggtacaccc ccgtgggccg ctccttcttc accgcgtccg aaggctgctc taaccctctt
     1981 ggcgggggcc gagaagtgtg gtttggcttc catcagtccg tccggccttc tctctggaaa
     2041 atgatgctga atattgatgt gtcagcaaca gcgttttaca aggcacagcc agtaatcgag
     2101 tttgtttgtg aagttttgga ttttaaaagt attgaagaac aacaaaaacc tctgacagat
     2161 tcccaaaggg taaagtttac caaagaaatt aaaggtctaa aggtggagat aacgcactgt
     2221 gggcagatga agaggaagta ccgtgtctgc aatgtgaccc ggcggcccgc cagtcaccaa
     2281 acattcccgc tgcagcagga gagcgggcag acggtggagt gcacggtggc ccagtatttc
     2341 aaggacaggc acaagttggt tctgcgctac ccccacctcc catgtttaca agtcggacag
     2401 gagcagaaac acacctacct tcccctggag gtctgtaaca ttgtggcagg acaaagatgt
     2461 attaaaaaat taacggacaa tcagacctca accatgatca gagcaactgc taggtcggcg
     2521 cccgatcggc aagaagagat tagcaaattg atgcgaagtg caagtttcaa cacagatcca
     2581 tacgtccgtg aatttggaat catggtcaaa gatgagatga cagacgtgac tgggcgggtg
     2641 ctgcagccgc cctccatcct ctacgggggc aggaataaag ctattgcgac ccctgtccag
     2701 ggcgtctggg acatgcggaa caagcagttc cacacgggca tcgagatcaa ggtgtgggcc
     2761 attgcgtgct tcgcccccca gcgccagtgc acggaagtcc atctgaagtc cttcacagag
     2821 cagctcagaa agatctcgag agacgctggc atgcccatcc agggccagcc gtgcttctgc
     2881 aaatacgcgc agggggcgga cagcgtggag cccatgttcc ggcacctgaa gaacacgtat
     2941 gcgggcctgc agctggtggt ggtcatcctg cccggcaaga cgcccgtgta cgccgaggtc
     3001 aagcgcgtgg gagacacggt gctggggatg gccacgcagt gcgtgcagat gaagaacgtg
     3061 cagaggacca cgccacagac cctgtccaac ctctgcctga agatcaacgt caagctggga
     3121 ggcgtgaaca acatcctgct gccccagggc aggccgccgg tgttccagca gcccgtcatc
     3181 tttctgggag cagacgtcac tcaccccccc gccggggatg ggaagaagcc ctccattgcc
     3241 gccgtggtgg gcagcatgga cgcccacccc aatcgctact gcgccaccgt gcgcgtgcag
     3301 cagcaccggc aggagatcat acaagacctg gccgccatgg tccgcgagct cctcatccag
     3361 ttctacaagt ccacgcgctt caagcccacc cgcatcatct tctaccgcga cggtgtctct
     3421 gaaggccagt tccagcaggt tctccaccac gagttgctgg ccatccgtga ggcctgtatc
     3481 aagctagaaa aagactacca gcccgggatc accttcatcg tggtgcagaa gaggcaccac
     3541 acccggctct tctgcactga caagaacgag cgggttggga aaagtggaaa cattccagca
     3601 ggcacgactg tggacacgaa aatcacccac cccaccgagt tcgacttcta cctgtgtagt
     3661 cacgctggca tccaggggac aagcaggcct tcgcactatc acgtcctctg ggacgacaat
     3721 cgtttctcct ctgatgagct gcagatccta acctaccagc tgtgtcacac ctacgtgcgc
     3781 tgcacacgct ccgtgtccat cccagcgcca gcatactacg ctcacctggt ggccttccgg
     3841 gccaggtacc acctggtgga taaggaacat gacagtgctg aaggaagcca tacctctggg
     3901 cagagtaacg ggcgagacca ccaagcactg gccaaggcgg tccaggttca ccaagacact
     3961 ctgcgcacca tgtactttgc ttgatctaga ccaagggcga attccagcac actggcggcc
     4021 gttactagtg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     4081 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4141 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4201 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4261 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     4321 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4381 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4441 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4501 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4561 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4621 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4681 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4741 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4801 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4861 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4921 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4981 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5041 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5161 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5221 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5281 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5341 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5401 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     5461 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5521 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5581 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5641 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5701 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5761 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     5821 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5881 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5941 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     6001 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6061 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6121 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6181 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     6241 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     6301 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     6361 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     6421 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     6481 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     6541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6601 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6661 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6721 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6781 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7141 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7201 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7261 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7321 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     7381 atagtaatca attacggggt cattagttca tagcccatat atg
//
