LOCUS       sequence-449336-        6692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of zebrafish Ago2 in zebrafish embryos..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  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 6692)
  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+-drAgo2
FEATURES             Location/Qualifiers
     source          1..6692
                     /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"
     polyA_signal    2744..2878
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2768..2787)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2822..2841
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2991..3011)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2991..3009)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3030..3046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3030..3046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3043..3065)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3054..3070
                     /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(3078..3084,3085..3102,3103..3108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3123..3144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3261..3278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3432..4020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3512..3531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4191..4982,4983..5051))
                     /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     4814..4833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5052..5156)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5182..5637)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5319..5340)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5531..5550
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5708..6692
                     /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 atccatgtat cccattggag cagctggtgc tactgagctg
      121 tttcagggac gtccgtcttc agggtcagat gtttctgctc ctgcgtctcc tccagcacca
      181 caagaatatg tcttcaaacc acctcagcgg ccagactttg gcacaatggg taggacaatc
      241 aagcttcagg ccaacttctt tgaaatggaa attcccaagt tggaggttta tcattatgaa
      301 atagacatca agcctgagaa atgcccacgg agagttaacc gtgaaatagt tgagcacatg
      361 gtccagcact ttaaaacgca gatctttgga gatcgaaagc ctgtgtatga tgggaggaag
      421 aatctctaca ctgccatgcc tttacccatt ggaagagaca aagtggagct ggaggtcacg
      481 attcctgggg agggaaaaga caggagcttt aaagtcgcta taaagtggat gtcctgtgtc
      541 agtctgcaag ctttgcatga agcactctct ggacggctac caaacatccc tttcgagacc
      601 attcaggctc tggatgttgt catgagacat ttgccctcta tgaggtacac cccagtcgga
      661 cgttcattct tcactccatc ggagggctgc tctaaccctc ttggtggagg aagagaagtt
      721 tggtttggtt ttcatcagtc tgttcggcca tcactctgga agatgatgct caacattgat
      781 gtgtctgcca ctgcgttcta caaagctcaa cccgtcattg aattcatgtg tgaggttttg
      841 gattttaaaa gcatcgaaga gcagcaaaag cccttaacag attcccaaag agtgaagttt
      901 accaaggaga tcaagggttt gaaggttgaa attactcact gtggacaaat gaagaggaag
      961 tacagagtgt gtaatgtgac gagaagacca gccagccatc aaacgtttcc tttgcagcaa
     1021 gagaatggtc agaccattga atgcactgtc gcccagtact tcaaggataa gtacaaactg
     1081 gtgcttcgat acccacatct cccatgttta caagttggtc aggagcagaa acacacctac
     1141 cttccgttgg aggtctgcaa cattgtagcc gggcagcgat gcatcaagaa actgacagac
     1201 aatcagacct ccactatgat acgtgccaca gccaggtctg cgcccgaccg tcaggacgag
     1261 atcagcaaac tgatgagaag tgccaacttc aatacagatc cttatgtgcg tgagtttgga
     1321 gtcatggtaa gagacgatat gactgaagtc aatggtcggg ttcttcaagc accgtcgatt
     1381 ctctatggtg gaaggaacaa agcaatagca accccagtcc agggcgtgtg ggacatgagg
     1441 aacaagcagt tccacacagg gattgaaatc aaagtttggg ccattgcctg ctttgcccca
     1501 cagaggcagt gcacagaact cctgctgaag gccttcactg accagcttcg taagatctca
     1561 cgtgatgctg ggatgcccat tcagggccag ccgtgctttt gcaaatacgc ccagggagca
     1621 gacagtgtgg agcccatgtt caaacacctc aagtacactt atcaagggtt acagttggtg
     1681 gtggtcatct tacccggcaa gactcctgtt tatgctgagg tgaagcgtgt tggagacact
     1741 gttctcggca tggccacgca gtgtgtccag gtgaagaatg tgcagaagac cacacctcaa
     1801 accctttcta acctctgcct caagattaac gtcaaactgg gtggagtcaa caacattctt
     1861 cttccacagg gcaggccctt ggtgttccag caaccagtta tctttttagg tgcagatgtg
     1921 actcatccac cagctggaga tggcaagaaa ccctcaattg ctgctgtcgt tggcagtatg
     1981 gatgcccacc caagcagata ctgtgccaca gtgcgggtcc agcagcaccg gcaggacatc
     2041 attcaggatc tggcaaccat ggtcagagag ttactcatcc agttctacaa atccacacgc
     2101 ttcaaaccaa cacgcatcat ctactacaga gacggcatct ctgaaggcca gtttaaccag
     2161 gttctacagc atgagctgct ggctattcgt gaggcctgca tcaagctgga gaaagattat
     2221 cagccgggta tcaccttcgt tgtagtacag aagagacacc acaccagact cttctgcatg
     2281 gacaggaacg aacgggttgg aaagagtgga aacatccctg ctggcaccac tgtcgacacc
     2341 aaaattactc atccattcga gtttgacttt tatctttgca gtcatgctgg aattcaggga
     2401 accagtaggc cgtcccacta tcatgtgctg tgggacgaca accatttcac ctctgacgag
     2461 ctgcaggtcc tcacctacca gctgtgccac acctatgttc gctgcacccg ctcggtctcc
     2521 attcctgcac cagcctatta cgcccacttg gtggcgtttc gcgcccgcta ccacttggtt
     2581 gacaaagaac atgacagtgc tgaaggcagt cacacatcag gtcagagtaa cggcagagat
     2641 caacaagccc tggccaaagc tgtgcagatc caccaggaca cactgcgcac catgtacttc
     2701 gcctgactcg agcctctaga actatagtga gtcgtattac gtagatccag acatgataag
     2761 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2821 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     2881 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     2941 attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     3001 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3061 ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc
     3121 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3181 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3241 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3301 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3361 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3421 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3481 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3541 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3601 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3661 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3721 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3781 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3841 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3901 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3961 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4021 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4081 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4141 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4201 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4261 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4321 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4381 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4441 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4501 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4561 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4621 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4681 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4741 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4801 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4861 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4921 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4981 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5041 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5101 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5161 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5221 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5281 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5341 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5401 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5461 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5521 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5581 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     5641 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5701 tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
     5761 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
     5821 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
     5881 ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
     5941 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
     6001 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
     6061 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
     6121 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
     6181 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
     6241 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
     6301 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
     6361 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
     6421 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
     6481 ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc
     6541 aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag
     6601 gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg
     6661 aggtctatat aagcaatgct cgtttaggga ac
//
