LOCUS       sequence-473518-        7287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  AUTHORS   Leung AK, Calabrese JM, Sharp PA
  TITLE     Quantitative analysis of Argonaute protein reveals 
            microRNA-dependent localization to stress granules.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Nov 28. 103(48):18125-30.
  PUBMED    17116888
REFERENCE   2  (bases 1 to 7287)
  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: "Proc Natl 
            Acad Sci U S A. 2006 Nov 28. 103(48):18125-30."
COMMENT     Sequence Label: EGFP-hAgo2
FEATURES             Location/Qualifiers
     source          1..7287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1457..2167,2168..2170,2171..2173))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1457..2167,2168..2170,2171..2173))
                     /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(1499..1520)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1848..1867
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2107..2128
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2170..2179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2218..2421)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2247..2267)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2422..2725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2900..3488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2980..2999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3817..3864
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3889..3908
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4096..4890)
                     /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             complement(4096..4890)
                     /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(4207..4226)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4817..4836
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4925..5282)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4939..5074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5091..5282
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5239..5259
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5284..5388)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5415..5870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5502..5521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5712..5733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5877..5998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5888..5907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5942..5961
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      6772..6781
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gctgctctgt gaaggacttc agatggactt ccgtgcactg gcgctggggg gcgaagcacg
       61 caatggccca caccttgatc tcgatgcccg tgtggaactg cttgttccgc atgtcccaga
      121 cgccctggac aggggtcgca atagctttat tcctgccccc gtagaggatg gagggcggct
      181 gcagcacccg cccagtcacg tctgtcatct catctttgac catgattcca aattcacgga
      241 cgtatggatc tgtgttgaaa cttgcacttc gcatcaattt gctaatctct tcttgccgat
      301 cgggcgccga cctagcagtc gctctgatca tggttgaggt ctgattgtcc gttaattttt
      361 taatacatct ttgtcctgcc acaatgttac agacctccag gggaaggtag gtgtgtttct
      421 gctcctgtcc gacttgtaaa catgggaggt gggggtagcg cagaaccaac ttgtgcctgt
      481 ccttgaaata ctgggccacc gtgcactcca ccgtctgccc gctctcctgc tgcagcggga
      541 atgtttggtg actggcgggc cgccgggtca cattgcagac acggtacttc ctcttcatct
      601 gcccacagtg cgttatctcc acctttagac ctttaatttc tttggtaaac tttacccttt
      661 gggaatctgt cagaggtttt tgttgttctt caatactttt aaaatccaaa acttcacaaa
      721 caaactcgat tactggctgt gccttgtaaa acgctgttgc tgacacatca atattcagca
      781 tcattttcca gagagaaggc cggacggact gatggaagcc aaaccacact tctcggcccc
      841 cgccaagagg gttagagcag ccttcggacg cggtgaagaa ggagcggccc acgggggtgt
      901 acctcatgga tggcaagtgc ctcatgacca cgtccagggc ctggatcgtc tcaaaaggga
      961 cgctgggcag ccgccctgaa agtgcatcgt gtaacgcctg caagctcacg caggacaccc
     1021 acttgatgga caccttgaag atgcgatcct tgccttctcc tggcagcgtg acctccagct
     1081 ccaccttgtc cctcccaatc ggaaggggca tggctgtgta tagattcttc ctgccgtcaa
     1141 acacgggctt ccgatcccca aagatctgtg ttttaaagtg ctggaccatg tgttccacga
     1201 tttccctgtt aactctcctc gggcacttct ctggcttgat atccaattca taatgataga
     1261 tgtcaatttt ggggatgtcc atttcgaaga aattggcctg taatttgatt gttctcccgg
     1321 aggtcccaaa gtcgggtcta ggtggaggct tgaaggcata tccttggatg gggggcggcg
     1381 gcgcaggagg tgcaagtgcg gggccggctc ccgagtacat ggaattcgaa gcttgagctc
     1441 gagatctgag tccggacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     1501 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     1561 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     1621 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     1681 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     1741 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     1801 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     1861 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     1921 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     1981 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     2041 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     2101 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     2161 ccttgctcac catggtggcg accggtagcg ctagcggatc tgacggttca ctaaaccagc
     2221 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     2281 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     2341 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     2401 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     2461 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     2521 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     2581 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     2641 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     2701 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     2761 cgtaattgat tactattaat aactaatgca tggcggtaat acggttatcc acagaatcag
     2821 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2881 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2941 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3001 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3061 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3121 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3241 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3301 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3361 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3421 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3481 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3541 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3601 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag
     3661 ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg
     3721 gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga
     3781 cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg
     3841 ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc
     3901 gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct
     3961 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     4021 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
     4081 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     4141 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     4201 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     4261 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     4321 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa
     4381 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     4441 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     4501 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     4561 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     4621 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     4681 cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt
     4741 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     4801 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     4861 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     4921 gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
     4981 aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     5041 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg
     5101 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     5161 ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     5221 agcctgggga ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc
     5281 aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5341 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5401 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5461 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5521 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5581 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5641 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5701 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5761 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5821 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt acgcgttaag
     5881 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     5941 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     6001 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     6061 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagttat ctagatccgg
     6121 tggatcctca agcaaagtac atggtgcgca gagtgtcttg gtgaacctgg accgccttgg
     6181 ccagtgcttg gtggtctcgc ccgttactct gcccagaggt atggcttcct tcagcactgt
     6241 catgttcctt atccaccagg tggtacctgg cccggaaggc caccaggtga gcgtagtatg
     6301 ctggcgctgg gatggacacg gagcgtgtgc agcgcacgta ggtgtgacac agctggtagg
     6361 ttaggatctg cagctcatca gaggagaaac gattgtcgtc ccagaggacg tgatagtgcg
     6421 aaggcctgct tgtcccctgg atgccagcgt gactacacag gtagaagtcg aactcggtgg
     6481 ggtgggtgat tttcgtgtcc acagtcgtgc ctgctggaat gtttccactt ttcccaaccc
     6541 gctcgttctt gtcagtgcag aagagccggg tgtggtgcct cttctgcacc acgatgaagg
     6601 tgatcccggg ctggtagtct ttttctagct tgatacaggc ctcacggatg gccagcaact
     6661 cgtggtggag aacctgctgg aactggcctt cagagacacc gtcgcggtag aagatgatgc
     6721 gggtgggctt gaagcgcgtg gacttgtaga actggatgag gagctcgcgg accatggcgg
     6781 ccaggtcttg tatgatctcc tgccggtgct gctgcacgcg cacggtggcg cagtagcgat
     6841 tggggtgggc gtccatgctg cccaccacgg cggcaatgga gggcttcttc ccatccccgg
     6901 cgggggggtg agtgacgtct gctcccagaa agatgacggg ctgctggaac accggcggcc
     6961 tgccctgggg cagcaggatg ttgttcacgc ctcccagctt gacgttgatc ttcaggcaga
     7021 ggttggacag ggtctgtggc gtggtcctct gcacgttctt catctgcacg cactgcgtgg
     7081 ccatccccag caccgtgtct cccacgcgct tgacctcggc gtacacgggc gtcttgccgg
     7141 gcaggatgac caccaccagc tgcaggcccg catacgtgtt cttcaggtgc cggaacatgg
     7201 gctccacgct gtccgccccc tgcgcgtatt tgcagaagca cggctggccc tggatgggca
     7261 tgccggcgtc tctcgagatc tttctga
//
