LOCUS       sequence-289332-        7366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAGO2-F2V2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7366)
  AUTHORS   Braun JE, Huntzinger E, Fauser M, Izaurralde E
  TITLE     GW182 proteins directly recruit cytoplasmic deadenylase complexes to
            miRNA targets.
  JOURNAL   Mol Cell. 2011 Oct 7;44(1):120-33.
  PUBMED    21981923
REFERENCE   2  (bases 1 to 7366)
  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: "Mol Cell.";
            date: "2011-10-7"; volume: "44(1)"; pages: "120-33"
COMMENT     Sequence Label: pT7-EGFP-C1-HsAGO2-F2V2_L
FEATURES             Location/Qualifiers
     source          1..7366
                     /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    4102..4223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4139..4158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4193..4212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4230..4685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4367..4388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4579..4598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4712..4816
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4818..5175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4818..5009
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4841..4861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5026..5161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5210..6004
                     /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             5210..6004
                     /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(5264..5283)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5874..5893
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6192..6211)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6236..6283
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6612..7200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7101..7120
                     /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 gagcgactgc 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 cgtcacagag
     2821 cagctcagaa agatctcgag agacgccggc atgcccatcc agggccagcc gtgcttctgc
     2881 aaatacgcgc agggggcgga cagcgtggag cccatggtcc 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 taactgatca taatcagcca taccacattt
     4021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4201 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     4261 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4321 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4381 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4441 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4501 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4561 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4621 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4681 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4741 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4801 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4921 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4981 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5041 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5101 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5161 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5221 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5281 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5341 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     5401 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5461 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5521 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5581 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5641 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5701 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5761 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5821 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5881 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5941 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6001 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6061 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6121 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6181 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     6241 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     6301 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     6361 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     6421 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     6481 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6541 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6721 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     6781 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7261 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     7321 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatg
//
