LOCUS       sequence-449343-        6698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human Ago2 in zebrafish embryos..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6698)
  AUTHORS   Chen GR, Sive H, Bartel DP
  TITLE     A Seed Mismatch Enhances Argonaute2-Catalyzed Cleavage and Partially
            Rescues Severely Impaired Cleavage Found in Fish.
  JOURNAL   Mol Cell. 2017 Dec 21;68(6):1095-1107.e5. doi: 
            10.1016/j.molcel.2017.11.032.
  PUBMED    29272705
REFERENCE   2  (bases 1 to 6698)
  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"; doi: 
            "10.1016/j.molcel.2017.11"; journalName: "Mol Cell"; date: 
            "2017-12-21- 21"; volume: "68"; issue: "6"; pages: "1095-1107.e5"
COMMENT     Sequence Label: pCS2+-hsAgo2
FEATURES             Location/Qualifiers
     source          1..6698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             93..113
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      2043..2052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2750..2884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2774..2793)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2828..2847
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2997..3017)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2997..3015)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3036..3052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3036..3052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3049..3071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3060..3076
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3084..3090,3091..3108,3109..3114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3129..3150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3267..3284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3438..4026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3518..3537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4197..4988,4989..5057))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4820..4839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5058..5162)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5188..5643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5325..5346)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5537..5556
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5714..6698
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccaacga ccgaaaacct gtattttcag ggcgccatgt
      121 actcgggagc cggccccgca cttgcacctc ctgcgccgcc gccccccatc caaggatatg
      181 ccttcaagcc tccacctaga cccgactttg ggacctccgg gagaacaatc aaattacagg
      241 ccaatttctt cgaaatggac atccccaaaa ttgacatcta tcattatgaa ttggatatca
      301 agccagagaa gtgcccgagg agagttaaca gggaaatcgt ggaacacatg gtccagcact
      361 ttaaaacaca gatctttggg gatcggaagc ccgtgtttga cggcaggaag aatctataca
      421 cagccatgcc ccttccgatt gggagggaca aggtggagct ggaggtcacg ctgccaggag
      481 aaggcaagga tcgcatcttc aaggtgtcca tcaagtgggt gtcctgcgtg agcttgcagg
      541 cgttacacga tgcactttca gggcggctgc ctagcgtccc ttttgagacg atccaggccc
      601 tggacgtggt catgaggcac ttgccatcca tgaggtacac ccccgtgggc cgctccttct
      661 tcaccgcgtc cgaaggctgc tctaaccctc ttggcggggg ccgagaagtg tggtttggct
      721 tccatcagtc cgtccggcct tctctctgga aaatgatgct gaatattgat gtgtcagcaa
      781 cagcgtttta caaggcacag ccagtaatcg agtttgtttg tgaagttttg gattttaaaa
      841 gtattgaaga acaacaaaaa cctctgacag attcccaaag ggtaaagttt accaaagaaa
      901 ttaaaggtct aaaggtggag ataacgcact gtgggcagat gaagaggaag taccgcgtct
      961 gcaatgtgac ccggcggccc gccagtcacc aaacattccc gctgcagcag gagagcgggc
     1021 agacggtgga gtgcacggtg gcccagtatt tcaaggacag gcacaagctt gttctgcgct
     1081 acccccacct cccatgttta caagtcggac aggagcagaa acacacctac cttcccctgg
     1141 aggtctgtaa cattgtggca ggacaaagat gtattaaaaa attaacggac aatcagacct
     1201 caaccatgat cagagcaact gctaggtcgg cgcccgatcg gcaagaagag attagcaaat
     1261 tgatgcgaag tgcaagtttc aacacagatc catacgtccg tgaatttgga atcatggtca
     1321 aagatgagat gacagacgtg actgggcggg tgctgcagcc gccctccatc ctctacgggg
     1381 gcaggaataa agctattgcg acccctgtcc agggcgtctg ggacatgcgg aacaagcagt
     1441 tccacacggg catcgagatc aaggtgtggg ccattgcgtg cttcgccccc cagcgccagt
     1501 gcacggaagt ccatctgaag tccttcacag agcagctcag aaagatctcg agagacgctg
     1561 gcatgcccat ccagggccag ccgtgcttct gcaaatacgc gcagggggcg gacagcgtgg
     1621 agcccatgtt ccggcacctg aagaacacgt atgcgggcct gcagctggtg gtggtcatcc
     1681 tgcccggcaa gacgcccgtg tacgccgagg tcaagcgcgt gggagacacg gtgctgggga
     1741 tggccacgca gtgcgtgcag atgaagaacg tgcagaggac cacgccacag accctgtcca
     1801 acctctgcct gaagatcaac gtcaagctgg gaggcgtgaa caacatcctg ctgccccagg
     1861 gcaggccgcc ggtgttccag cagcccgtca tctttctggg agcagacgtc actcaccccc
     1921 ccgccgggga tgggaagaag ccctccattg ccgccgtggt gggcagcatg gacgcccacc
     1981 ccaatcgcta ctgcgccacc gtgcgcgtgc agcagcaccg gcaggagatc atacaagacc
     2041 tggccgccat ggtccgcgag ctcctcatcc agttctacaa gtccacgcgc ttcaagccca
     2101 cccgcatcat cttctaccgc gacggtgtct ctgaaggcca gttccagcag gttctccacc
     2161 acgagttgct ggccatccgt gaggcctgta tcaagctaga aaaagactac cagcccggga
     2221 tcaccttcat cgtggtgcag aagaggcacc acacccggct cttctgcact gacaagaacg
     2281 agcgggttgg gaaaagtgga aacattccag caggcacgac tgtggacacg aaaatcaccc
     2341 accccaccga gttcgacttc tacctgtgta gtcacgctgg catccagggg acaagcaggc
     2401 cttcgcacta tcacgtcctc tgggacgaca atcgtttctc ctctgatgag ctgcagatcc
     2461 taacctacca gctgtgtcac acctacgtgc gctgcacacg ctccgtgtcc atcccagcgc
     2521 cagcatacta cgctcacctg gtggccttcc gggccaggta ccacctggtg gataaggaac
     2581 atgacagtgc tgaaggaagc catacctctg ggcagagtaa cgggcgagac caccaagcac
     2641 tggccaaggc ggtccaggtt caccaagaca ctctgcgcac catgtacttt gcttgatagg
     2701 atcgaatcgg actttcgaat tctagaacta tagtgagtcg tattacgtag atccagacat
     2761 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2821 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2881 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2941 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     3001 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3061 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     3121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4141 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5161 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     5221 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     5281 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     5341 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     5401 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5461 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5521 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5581 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5641 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5701 cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa
     5761 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
     5821 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     5881 tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     5941 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     6001 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     6061 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     6121 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     6181 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     6241 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     6301 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     6361 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
     6421 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
     6481 taacttggca agtacattac tattggaagt acgccagggt acattggcag tactcccatt
     6541 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
     6601 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
     6661 aatgggaggt ctatataagc aatgctcgtt tagggaac
//
