LOCUS       sequence-174835-        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"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     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"
     regulatory      3292..3301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4075..4196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4112..4131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4166..4185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4203..4658)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4340..4361)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4552..4571
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4685..4789
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4791..5148
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4791..4982
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4814..4834)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4999..5134
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5183..5977
                     /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             5183..5977
                     /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(5237..5256)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5847..5866
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6165..6184)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6209..6256
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6585..7173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7074..7093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      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 attccatgta ctcgggagcc
     1381 ggccccgcac ttgcacctcc tgcgccgccg ccccccatcc aaggatatgc cttcaagcct
     1441 ccacctagac ccgactttgg gacctccggg agaacaatca aattacaggc caatttcttc
     1501 gaaatggaca tccccaaaat tgacatctat cattatgaat tggatatcaa gccagagaag
     1561 tgcccgagga gagttaacag ggaaatcgtg gaacacatgg tccagcactt taaaacacag
     1621 atctttgggg atcggaagcc cgtgtttgac ggcaggaaga atctatacac agccatgccc
     1681 cttccgattg ggagggacaa ggtggagctg gaggtcacgc tgccaggaga aggcaaggat
     1741 cgcatcttca aggtgtccat caagtgggtg tcctgcgtga gcttgcaggc gttacacgat
     1801 gcactttcag ggcggctgcc cagcgtccct tttgagacga tccaggccct ggacgtggtc
     1861 atgaggcact tgccatccat gaggtacacc cccgtgggcc gctccttctt caccgcgtcc
     1921 gaaggctgct ctaaccctct tggcgggggc cgagaagtgt ggtttggctt ccatcagtcc
     1981 gtccggcctt ctctctggaa aatgatgctg aatattgatg tgtcagcaac agcgttttac
     2041 aaggcacagc cagtaatcga gtttgtttgt gaagttttgg attttaaaag tattgaagaa
     2101 caacaaaaac ctctgacaga ttcccaaagg gtaaagttta ccaaagaaat taaaggtcta
     2161 aaggtggaga taacgcactg tgggcagatg aagaggaagt accgtgtctg caatgtgacc
     2221 cggcggcccg ccagtcacca aacattcccg ctgcagcagg agagcgggca gacggtggag
     2281 tgcacggtgg cccagtattt caaggacagg cacaagttgg ttctgcgcta cccccacctc
     2341 ccatgtttac aagtcggaca ggagcagaaa cacacctacc ttcccctgga ggtctgtaac
     2401 attgtggcag gacaaagatg tattaaaaaa ttaacggaca atcagacctc aaccatgatc
     2461 agagcgactg ctaggtcggc gcccgatcgg caagaagaga ttagcaaatt gatgcgaagt
     2521 gcaagtttca acacagatcc atacgtccgt gaatttggaa tcatggtcaa agatgagatg
     2581 acagacgtga ctgggcgggt gctgcagccg ccctccatcc tctacggggg caggaataaa
     2641 gctattgcga cccctgtcca gggcgtctgg gacatgcgga acaagcagtt ccacacgggc
     2701 atcgagatca aggtgtgggc cattgcgtgc ttcgcccccc agcgccagtg cacggaagtc
     2761 catctgaagt ccttcacaga gcagctcaga aagatctcga gagacgccgg catgcccatc
     2821 cagggccagc cgtgcttctg caaatacgcg cagggggcgg acagcgtgga gcccatgttc
     2881 cggcacctga agaacacgta tgcgggcctg cagctggtgg tggtcatcct gcccggcaag
     2941 acgcccgtgt acgccgaggt caagcgcgtg ggagacacgg tgctggggat ggccacgcag
     3001 tgcgtgcaga tgaagaacgt gcagaggacc acgccacaga ccctgtccaa cctctgcctg
     3061 aagatcaacg tcaagctggg aggcgtgaac aacatcctgc tgccccaggg caggccgccg
     3121 gtgttccagc agcccgtcat ctttctggga gcagacgtca ctcacccccc cgccggggat
     3181 gggaagaagc cctccattgc cgccgtggtg ggcagcatgg acgcccaccc caatcgctac
     3241 tgcgccaccg tgcgcgtgca gcagcaccgg caggagatca tacaagacct ggccgccatg
     3301 gtccgcgagc tcctcatcca gttctacaag tccacgcgct tcaagcccac ccgcatcatc
     3361 ttctaccgcg acggtgtctc tgaaggccag ttccagcagg ttctccacca cgagttgctg
     3421 gccatccgtg aggcctgtat caagctagaa aaagactacc agcccgggat caccttcatc
     3481 gtggtgcaga agaggcacca cacccggctc ttctgcactg acaagaacga gcgggttggg
     3541 aaaagtggaa acattccagc aggcacgact gtggacacga aaatcaccca ccccaccgag
     3601 ttcgacttct acctgtgtag tcacgctggc atccagggga caagcaggcc ttcgcactat
     3661 cacgtcctct gggacgacaa tcgtttctcc tctgatgagc tgcagatcct aacctaccag
     3721 ctgtgtcaca cctacgtgcg ctgcacacgc tccgtgtcca tcccagcgcc agcatactac
     3781 gctcacctgg tggccttccg ggccaggtac cacctggtgg ataaggaaca tgacagtgct
     3841 gaaggaagcc atacctctgg gcagagtaac gggcgagacc accaagcact ggccaaggcg
     3901 gtccaggttc accaagacac tctgcgcacc atgtactttg cttgaggatc caccggatct
     3961 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4021 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     4081 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4141 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     4201 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4261 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4321 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4381 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4441 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4501 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     4561 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4621 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     4681 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4741 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4801 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4861 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4921 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4981 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5041 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5101 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     5161 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5221 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5281 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5341 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     5401 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5461 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5521 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5581 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5641 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5701 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5761 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5821 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5881 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5941 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     6001 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     6061 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     6121 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     6181 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     6241 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     6301 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     6361 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     6421 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6481 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6541 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6601 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6661 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6721 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6781 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6841 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6901 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6961 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7021 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7081 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7141 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7201 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7261 gtggataacc gtattaccgc catgcat
//
