LOCUS       sequence-288673-        7798 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 7798)
  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 7798)
  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..7798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2588..4243
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVA
                     PANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQ
                     SMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSH
                     ARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDY
                     KIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQG
                     YGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIM
                     SGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESI
                     LLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGV
                     VFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2657..2675)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    4254..4278
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5073..5097)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      complement(5981..6569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6061..6080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6740..7531,7532..7600))
                     /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     7363..7382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7601..7705)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
     2581 taccaacatg gccgaagacg ccaaaaacat aaagaaaggc ccggcaccat tctatccact
     2641 agaggatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     2701 tggaacaatt gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt
     2761 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     2821 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     2881 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     2941 tatgaacatt tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt
     3001 gaacgtgcaa aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga
     3061 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     3121 tgaatacgat tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa
     3181 ctcctctgga tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt
     3241 cagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     3301 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     3361 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct
     3421 tcaggattac aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa
     3481 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc
     3541 acctctttcg aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg
     3601 acaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     3661 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     3721 taccgggaaa acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat
     3781 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     3841 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg
     3901 cttgaagtct ttaattaaat acaaagggta tcaggtggcc cccgctgaat tggaatcgat
     3961 attgttacaa caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc
     4021 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     4081 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     4141 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     4201 gatcctcata aaggccaaga agggcggaaa gtccaaattg taagaattat caaacaagtt
     4261 tgtacaaaaa agcaggcttc tctagattta gctagtttag gaacttttgg gagtgcaatg
     4321 aacttacaac gaaagccaaa gcaaccaacc aaagatatat aactaatact agtcagtcat
     4381 atatccatct actcctttgc tccgacatca tctcatgaca tcattcatca cccatatcat
     4441 ccttcattct tcctactact aatcatccca tagcttttca cactttcata ctaatttcct
     4501 catgactcac tctcattctg tatcatcttt gcattttttg caaatcattt tccagttaat
     4561 attctaccca aaattcgacc ctttaacaac tttttcaatc tggaaatttc tcgtcattct
     4621 agtcacaagt tcttgcattc cttctagtca cttagggtaa atcatctcta taagtcagct
     4681 tcagttcgat cttactaagt ttgtacaaac atatatcgtt cttcaatttc ggatacagct
     4741 ttagtcatat tcaggttata taccgaaacc aaaattcagg caaaattgtg cagtaatagt
     4801 gtttaagttc gaatcaaacc aaaaagcaaa ccaaatccta gtttaaagca aattcaagcc
     4861 cagtgtttgt gaaccaaagt cgagtgtgag ctctagtcat tttagccacc taatcaagcg
     4921 gaatcagctt gttctcagtt tagtttagtt agttaagtta ggcatagaaa cggacattgg
     4981 accagcgtct cactaccagt caaagtattg aacagtacaa cctagctata agggccacag
     5041 gcttcagaac cggcggttgt gtacttaaag cgacccagct ttcttgtaca aagtggttcg
     5101 ataattcgct agcactagag tgtacatcct gtgtaccctt attaagcaat cggtcctttt
     5161 caggaccacc attcagtttt taaaaggagg aagattttca aaaagtattt aattttattg
     5221 gtcgggactc ccaaaaaata tgttaaacta aaaataaatg tctgtcgttc gtactctttc
     5281 aaatagaatg tgtatttcta acatggattt gtacagactg tacaaatggt ttcagacacc
     5341 gcatgtacag catcctcgaa acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     5401 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     5461 tgatcagtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     5521 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     5581 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     5641 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     5701 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg
     5761 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta
     5821 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc
     5881 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagccagc
     5941 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6001 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6061 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6121 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     6181 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6241 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6301 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6361 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6421 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6481 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6541 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6601 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6661 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6721 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6781 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6841 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6901 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6961 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7021 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     7081 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7141 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7201 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7261 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7321 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     7381 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7441 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7501 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7561 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7621 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7681 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     7741 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagc cctttcgt
//
