LOCUS       sequence-288471-        9158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmStaufen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9158)
  AUTHORS   Behm-Ansmant I, Rehwinkel J, Doerks T, Stark A, Bork P, Izaurralde E
  TITLE     mRNA degradation by miRNAs and GW182 requires both CCR4:NOT 
            deadenylase and DCP1:DCP2 decapping complexes.
  JOURNAL   Genes Dev. 2006 Jul 15. 20(14):1885-98.
  PUBMED    16815998
REFERENCE   2  (bases 1 to 9158)
  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: "Genes Dev. 
            2006 Jul 15. 20(14):1885-98."
COMMENT     Sequence Label: pAc5.1B-EGFP-DmStaufen-V5His6_C
FEATURES             Location/Qualifiers
     source          1..9158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /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(2633..2654)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2894..2913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             5914..5922
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             6443..6484
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6494..6511
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6534..6551)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6584..6718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6608..6627)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6662..6681
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7340..7928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7420..7439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8099..8890,8891..8959))
                     /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     8722..8741
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8960..9064)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9132..9150
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
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 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3301 caagccaagc ttgaattctg cagatatcat gcagcacaac gttcatgccg cccggccagc
     3361 gccccacatc cgagctgcgc accatcacag ccacagccat gcccacatgc atctgcatcc
     3421 cggcatggag cagcacctgg gccccagtct tcagcagcaa cagcagccgc caccgccgcc
     3481 ccagcagcct ccacatcgcg acctccacgc acgtctgaat catcaccatc tccacgctca
     3541 gcagcagcag cagcaacaga cgtcgtcgaa ccaagccgcc gccgtcgctg cggctggggc
     3601 tgcctatcac cacggcaaca tcaacagcaa cagcggcagc aacatctcca gcaacagcaa
     3661 ccagatgcag aagatccgcc aacagcacca gcacttgagt agcagcaacg gccttcttgg
     3721 caatcaacct cctggtccgc cgccacaggc cttcaatcca ttggcgggca atccggcagc
     3781 tctcgcctac aatcagcttc ctccgcatcc gccgcaccac atggccgccc atctgggcag
     3841 ctatgccgct ccaccgcctc actattacat gagccaggcg aagcccgcga aatataatca
     3901 ctatggcagc aatgcaaaca gcaacagcgg cagcaacaac agcaacagca actatgcccc
     3961 caaggccata ctgcagaata cgtataggaa tcagaaggtg gtggttccac cagtggtgca
     4021 ggaagtaacc ccggtgccgg aaccgccagt gaccaccaat aatgccacca ccaacagcac
     4081 cagcaacagc acagtcatag ctagcgagcc tgtaacccaa gaggatactt ctcagaagcc
     4141 ggagactaga caggagccag cttcagcgga cgatcatgtc agcacgggga atatcgatgc
     4201 tacgggagca ctgtccaacg aggacacgag cagttcagga cgcgggggca aggacaagac
     4261 cccaatgtgc ctggtcaatg agttggccag gtacaacaag atcacgcatc aatatcggtt
     4321 gactgaagaa cgaggacctg cccactgcaa gaccttcacg gtgacactga tgctgggcga
     4381 tgaggagtac tcggcggatg gttttaagat taagaaagcg caacatttgg cggctagtaa
     4441 ggctatcgag gaaaccatgt acaagcatcc gccgccgaag attcgccgca gcgaagaagg
     4501 cggtcccatg cgcacacaca ttacgccgac ggtggaattg aatgccctgg ccatgaagtt
     4561 gggacagcgc accttctacc tgctggatcc cactcagatt cctcccacgg attccatagt
     4621 gccgccggag tttgccggcg gtcatttact aactgcacca ggaccaggga tgccgcaacc
     4681 acctccacca cctgcttacg ctctgcgcca gcggcttggc aatggatttg tgcccattcc
     4741 atcgcagccg atgcatccgc atttctttca tggacctggt cagcgaccgt ttcctccgaa
     4801 gtttccatca cgctttgcgc tgccaccgcc tctcggagct catgtacatc atggaccgaa
     4861 tggcccattt ccatccgtgc ctacgccacc cagcaaaatc accctgtttg tgggcaagca
     4921 gaagtttgtg ggcatcggac gcacactgca gcaagcgaaa catgacgctg cggcgagagc
     4981 gctgcaggtt ctcaagacac aggccatatc tgcctctgag gaggctctcg aggattcgat
     5041 ggatgagggt gacaagaagt cgcccatctc gcaggttcac gagatcggaa ttaagcgcaa
     5101 catgaccgtt cattttaagg tgttgcgcga ggaggggccg gcgcacatga agaactttat
     5161 aacagcctgc attgtgggat ctattgtcac cgaaggagaa ggaaatggca aaaaagtgtc
     5221 caagaagcgt gccgcggaaa agatgctcgt cgagttgcaa aagctgccgc cccttacgcc
     5281 caccaagcag acgccattga agcggattaa ggttaagaca ccaggcaaaa gtggtgcagc
     5341 tgcccgagaa ggttcggttg tgtcgggcac tgatggtact atgcaaacgg gaaagccgga
     5401 gagacgtaaa cgcctcaatc cgccgaagga caagctgata gacatggacg atgcggataa
     5461 tcccattaca aagctaattc aactgcagca gacgcgtaag gaaaaggaac ccattttcga
     5521 gctgatagcc aaaaacggca atgaaaccgc tcgacgacgg gaattcgtta tggaggtctc
     5581 cgccagcggg agcacagccc gtggaacagg caacagcaag aaattggcaa agcgaaatgc
     5641 tgcacaggct ctatttgaac tgctggaagc tgttcaagtc acaccaacca acgaaacaca
     5701 gtcatcggag gaatgctgca cttcggcaac aatgtccgcc gttacagcgc ccgcagtgga
     5761 ggcaactgcg gaaggcaaag tgcccatggt agcaacgcca gtgggtccca tgccaggcat
     5821 tttaatactg cgccagaaca agaaaccagc aaagaagagg gatcaaattg taattgtcaa
     5881 gtctaacgtg gaatccaaag aggaggaggc caataaggag gtggcagttg cagctgagga
     5941 gaatagcaac aacagtgcaa actctggcga cagcagcaac agcagcagcg gcgattcgca
     6001 agccacagag gcggctagtg agagtgcact gaatacctca accggcagca atacgagtgg
     6061 tgtgagtagc aacagcagca atgttggagc caacacggat ggtaacaacc atgccgagag
     6121 taaaaacaac acggagagta gcagcaacag tacgagtaac acacaaagcg ctggagtgca
     6181 catgaaggag cagctcttgt accttagtaa actgctggac tttgaggtca acttctcgga
     6241 ctatccgaaa ggcaatcaca acgagttcct gaccatcgtg acattgtcca cacatccgcc
     6301 gcagatttgc catggcgttg gcaagagctc cgaggagtcg cagaatgatg ctgcaagcaa
     6361 tgccttgaaa atcctcagca agctggggct caacaatgcc atgaaaggcg gccgctcgag
     6421 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     6481 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     6541 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     6601 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     6661 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     6721 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     6781 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga ccgatgccct
     6841 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     6901 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     6961 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     7021 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     7081 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
     7141 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
     7201 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     7261 gacagcttca aggatcgctc gcggctctta ccagccagca aaaggccagg aaccgtaaaa
     7321 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7381 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7441 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7501 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     7561 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7621 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7681 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7741 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7801 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7861 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7921 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7981 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8041 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8101 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8161 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8221 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8281 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8341 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8401 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8461 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8521 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8581 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8641 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8701 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8761 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8821 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8881 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8941 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9001 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9061 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     9121 taacctataa aaataggcgt atcacgaggc cctttcgt
//
