LOCUS       sequence-355100-        7775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of Nerfin-3UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7775)
  AUTHORS   Eulalio A, Huntzinger E, Izaurralde E
  TITLE     GW182 interaction with Argonaute is essential for miRNA-mediated 
            translational repression and mRNA decay.
  JOURNAL   Nat Struct Mol Biol. 2008 Apr . 15(4):346-53.
  PUBMED    18345015
REFERENCE   2  (bases 1 to 7775)
  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: "Nat Struct 
            Mol Biol. 2008 Apr . 15(4):346-53."
COMMENT     Sequence Label: pAc5.1C-Fluc-nerfin-HSL_E
FEATURES             Location/Qualifiers
     source          1..7775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..2510
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2530..4185
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVA
                     PANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQ
                     SMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSH
                     ARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDY
                     KIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQG
                     YGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIM
                     SGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESI
                     LLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGV
                     VFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2599..2617)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    4196..4220
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5015..5039)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      complement(5902..6490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5982..6001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6661..7452,7453..7521))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7284..7303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7522..7626)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7694..7712
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catgaatggc atcaactctg aatcaaatct ttgcagatgc acctacttct catttccact
       61 gtcacatcat ttttccagat ctcgctgcct gttatgtggc ccacaaacca agacacgttt
      121 tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacataatga atatgtacac
      181 attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga gtaggagaat gcggaggaga
      241 aggagaacga gctgatctag tatctctcca caatccaatg ccaactgacc aactggccat
      301 attcggagca atttgaagcc aatttccatc gcctggcgat cgctccattc ttggctatat
      361 gtttttcacc gttacccggg gccattttca aagactcgtc ggcaagataa gattgtgtca
      421 ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag
      481 tattttgaag aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac
      541 aaaaacatgt tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt
      601 atgtatatac ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac
      661 tgatgatatt ttaagatgat gccagaccaa aaggcttgaa tttctgcgtc ttttgccgaa
      721 cgcagtgcat gtgcaattgt tgttttttgg aatattcaat tttcggactg tccgctttga
      781 tttcagtttc ttggcttatt caaaaagcaa agtaaagcca aaaaagcgag atggcaatac
      841 caaatgcggc aaaacggtag tggaaggaaa ggggtgcggg gcagcggaag gaagggtggg
      901 gcggggcgtg gcggggtctg tggctgggcg cgacgtcacc gacgttggag ccactccttt
      961 gaccatgtgt gcgtgtgtgt attattcgtg tctcgccact cgccggttgt ttttttcttt
     1021 ttatgctgcg ctctctctag cgccatctcg cttacgcatg ctcaacgcac cgcatgttgc
     1081 cgtttccttt tatgcgtcat tttggctcga aataggcaat tatttaaaca aagattagtc
     1141 aacgaaaacg ctaaaataaa taagtctaca atatggttac ttattgccat gtgtgtgcag
     1201 ccaacgatag caacaaaagc aacaacacag gtggctttcc ctctttcact ttttgtttgc
     1261 aagccgcgtg cgagcaagac ggcacgaccg gcaaacgcaa ttacgctgac aaagagcaga
     1321 cgaagttttg gcgaaaaaca tcaaggcgcc tgatacgaat gcatttgcaa taacaattgc
     1381 gatatttaat attgtttatg aagctgtttg acttcaaaac acacaaaaaa aaaaataaaa
     1441 caaattattt gaaagagaat taggaatcgg acgcttatcg ttagggtaac aacaagaaat
     1501 gcttactgag tcacagcctc tggaaaactg ccgcaagcca gagagagaga gaaaaagagg
     1561 gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg tctctgttgc
     1621 gcaaacgcat agactgcact gaaaaaatcg attacctatt ttttatgaat gaatatttgc
     1681 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1741 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1801 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1861 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgaatgcc ctactagaag
     1921 atgtgtgttg cacaaaatgt ccctggaata accaatttga agtgcagata gcagtaaacg
     1981 taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca ttactaataa
     2041 acccactata aacacatgta catatgtatg ttttggcata caatgagtag ttggggaaaa
     2101 aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag tatcgaatat
     2161 gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa gtacactctt
     2221 catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga aattctccgt
     2281 aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt ttgaacgcga
     2341 cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc tttataaaac
     2401 gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt gtggatactc
     2461 ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac cccggatcgg
     2521 ggtaccaaca tggccgaaga cgccaaaaac ataaagaaag gcccggcacc attctatcca
     2581 ctagaggatg gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt
     2641 cctggaacaa ttgcttttac agatgcacat atcgaggtga acatcacgta cgcggaatac
     2701 ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac
     2761 agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta
     2821 tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac
     2881 agtatgaaca tttcgcagcc taccgtagtg tttgtttcca aaaaggggtt gcaaaaaatt
     2941 ttgaacgtgc aaaaaaaatt accaataatc cagaaaatta ttatcatgga ttctaaaacg
     3001 gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt
     3061 aatgaatacg attttgtacc agagtccttt gatcgtgaca aaacaattgc actgataatg
     3121 aactcctctg gatctactgg gttacctaag ggtgtggccc ttccgcatag aactgcctgc
     3181 gtcagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg
     3241 attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg
     3301 atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tttacgatcc
     3361 cttcaggatt acaaaattca aagtgcgttg ctagtaccaa ccctattttc attcttcgcc
     3421 aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc ttctgggggc
     3481 gcacctcttt cgaaagaagt cggggaagcg gttgcaaaac gcttccatct tccagggata
     3541 cgacaaggat atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat
     3601 gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg
     3661 gataccggga aaacgctggg cgttaatcag agaggcgaat tatgtgtcag aggacctatg
     3721 attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga
     3781 tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt catagttgac
     3841 cgcttgaagt ctttaattaa atacaaaggg tatcaggtgg cccccgctga attggaatcg
     3901 atattgttac aacaccccaa catcttcgac gcgggcgtgg caggtcttcc cgacgatgac
     3961 gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa
     4021 gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt
     4081 gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga
     4141 gagatcctca taaaggccaa gaagggcgga aagtccaaat tgtaagaatt atcaaacaag
     4201 tttgtacaaa aaagcaggct tctctagatt tagctagttt aggaactttt gggagtgcaa
     4261 tgaacttaca acgaaagcca aagcaaccaa ccaaagatat ataactaata ctagtcagtc
     4321 atatatcctt ctactccttt gctccgacat catctcatga catcattcat cacccatatc
     4381 atccttcatt cttcctacta ctaatcatcc catagctttt cacactttca tactaatttc
     4441 ctcatgactc actctcattc tgtatcatct ttgcattttt tgcaaatcat tttccagtta
     4501 atattctacc caaaattcga ccctttaaca actttttcaa tctggaaatt tctcgtcatt
     4561 ctagtcacaa gttcttgcat tccttctagt cacttagggt aaatcatctc tataagtcag
     4621 cttcagttcg atcttactaa gtttgtacaa acatatatcg ttcttcaatt tcggatacag
     4681 ctttagtcat attcaggtta tataccgaaa ccaaaattca ggcaaaattg tgcagtaata
     4741 gtgtttaagt tcgaatcaaa ccaaaaagca aaccaaatcc tagtttaaag caaattcaag
     4801 cccagtgttt gtgaaccaaa gtcgagtgtg agctctagtc attttagcca cctaatcaag
     4861 cggaatcagc ttgttctcag tttagtttag ttagtttagt taggcataga aacggacatt
     4921 ggaccagcgt ctcactacca gtcaaagtat tgaacagtac aacctagcta taagggccac
     4981 aggcttcaga accggcggtt gtgtacttaa agcgacccag ctttcttgta caaagtggtt
     5041 cgataattcg ctagcactag agtgtacatc ctgtgtaccc ttattaagca atcggtcctt
     5101 ttcaggacca ccattcagtt tttaaaagga ggaagatttt caaaaagtat ttaattttat
     5161 tggtcgggac tcccaaaaaa tatgttaaac taaaaataaa tgtctgtcgt tcgtactctt
     5221 tcaaatagaa tgtgtatttc taacatggat ttgtacagac tgtacaaatg gtttcagaca
     5281 ccgcatgtac agcatcctcg aaacaacaac aattgcattc attttatgtt tcaggttcag
     5341 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     5401 tatgatcagt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     5461 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     5521 acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac
     5581 gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt
     5641 cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc
     5701 cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat
     5761 tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag
     5821 gcaggtagat gacgaccatc agggacagct tcaaggccag caaaaggcca ggaaccgtaa
     5881 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5941 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6001 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6061 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6121 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6181 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6241 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6301 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6361 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6421 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6481 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6541 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6601 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6661 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6721 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6781 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6841 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6901 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6961 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7021 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7081 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7141 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7201 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7261 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     7321 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7381 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7441 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7501 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7561 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7621 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     7681 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat taattctata
     7741 ttctaaaaac acaaatgata cttctaaaaa aaaat
//
