LOCUS       sequence-289833-        8748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of CeAIN2_254-400-DmGW182_635-1384-F961A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8748)
  AUTHORS   Huntzinger E, Kuzuoglu-Ozturk D, Braun JE, Eulalio A, Wohlbold L, 
            Izaurralde E
  TITLE     The interactions of GW182 proteins with PABP and deadenylases are 
            required for both translational repression and degradation of miRNA 
            targets.
  JOURNAL   Nucleic Acids Res. 2013 Jan;41(2):978-94. doi: 10.1093/nar/gks1078. 
            Epub 2012 Nov 21.
  PUBMED    23172285
REFERENCE   2  (bases 1 to 8748)
  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: 
            "10.1093/nar/gks1078"; journalName: "Nucleic Acids Res"; date: 
            "2013-01"; volume: "41"; issue: "2"; pages: "978-94"
COMMENT     Sequence Label: pAc5.1B-EGFP-CeAIN2_254-400-DmGW182_635-1384-F...
FEATURES             Location/Qualifiers
     source          1..8748
                     /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"
     CDS             6033..6074
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6084..6101
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6124..6141)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6174..6308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6198..6217)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6252..6271
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6930..7518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7010..7029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7689..8480,8481..8549))
                     /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     8312..8331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8550..8654)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8722..8740
                     /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 ttaatggatg attattcaat gtggactgat gagaatgatg aagacgcgaa
     3361 aaagaaaaaa tcgttgagag ataagggatt gagtggatgg ggtgatgctg atgtatcaaa
     3421 cagtaaacca attcgccgtt ggattgtgcc agaaggccaa gaagaagact ttgaaacggc
     3481 aatggaacga tgtccaagtc atttgaaaaa gaaagcattg aatgaagaag ctcttcgtcg
     3541 acgtttaaca tctgaaaatc cacaaatagt ggcacaagct caacaacaag ccgaagaaga
     3601 ggcgaatgct atgatgagga ttggacgaag acccattgtt cccagtggat ggggtgatat
     3661 tccatctgaa attagtactg acaagtcaga attcgatcaa agctcgtcac gtggttggga
     3721 tgaaggaagt ataagcggtg gtggaagttg cggccgccat aataacctta tgaataacat
     3781 gggaggtacc gctgttactg gaggtaacaa caatacaaac atgacagctt tacaggtgca
     3841 aaagtattta aatcaagggc aacatggtgt cgctgttgga ccgcaagccg ttggtaattc
     3901 ttcagcagta tctgtcggat ttggtcagaa cacgtctaac gcagcagtgg caggagcagc
     3961 ctctgcaaat atagcagcaa atacaaacaa ccaaccgtct ggtcagcaaa ttcgcatgct
     4021 aggccagcaa attcagttgg ccattcatag tggtttcata tctagtcaga tattgactca
     4081 accgctaact caaacaaccc ttaacctttt aaaccaactt cttagcaata ttaagcatct
     4141 ccaggctgca cagcaatccc ttacccgcgg gggaaatgtc aatccaatgg cagtgaatgt
     4201 ggctatatct aaatacaagc agcaaatcca gaatttacag aaccagataa gtgcacaaca
     4261 ggctgtgtat gtaaaacagc aaaatatgca accaacttca caacaacaac agccccaaca
     4321 acagcaactt ccttctgttc atctaagtaa ctcaggcaac gactatttaa gaggtcacga
     4381 tgcaataaat aatttgcaaa gcaacttttc tgagctcaat attaataagc caagtggata
     4441 tcaaggagcg tccaatcaac aatcccgatt aaatcagtgg aagcttccag tattagataa
     4501 ggagatcaac tctgacagta cggaattttc tcgtgcccca ggtgcaacga aacaaaattt
     4561 gacggccaac acaagcaaca taaactcttt gggtcttcaa aacgatagta catggtcaac
     4621 tggacgcagt attggtgacg gttggcctga tccctcatct gataacgaga ataaagactg
     4681 gtctgttgct cagccaactt cagcagcaac tgcttacact gatctggtcc aagaggctga
     4741 gccaggcaag ccatggaagg gttcacagat caaaagcata gaagatgatc ccagcattac
     4801 accaggaagc gttgctagat ctccattgtc tattaattcg acgccaaaag atgctgacat
     4861 atttgccaat accggtaaaa attcaccgac tgatttaccg ccactaagtt tatcgtcgtc
     4921 tacatggagt tttaatccaa accaaaatta tccgagtcac agttggtctg acaatagtca
     4981 acaatgtacc gccacttcgg agctttggac aagcccgcta aataaatcat cgtctcgagg
     5041 tcccccgcca ggattgactg ccaattcaaa taaatctgca aatagtaatg cgtcaacgcc
     5101 aacaactatt accggaggtg cgaatggatg gttacagcct cgaagtggcg gtgttcaaac
     5161 cacaaacact aattggacag gtggtaacac cacttggggc tccagttggt tgcttttgaa
     5221 aaatctaaca gcacagattg atggtcctac tttgcgtaca ctgtgtatgc agcatgggcc
     5281 ccttgtcagc tttcacccgt atttgaacca aggaattgcc ttatgtaaat atactactcg
     5341 tgaggaggcg aacaaggcgc aaatggcgtt aaacaactgt gtcctcgcca acaccacaat
     5401 atttgctgaa tctcccagcg agaacgaggt gcaaagcatt atgcagcact taccacaaac
     5461 tccttcctct acaagctcta gtggaactag tggtggcaac gtcggaggcg tcggcacttc
     5521 agccaataat gcaaacagtg gttctgcagc ttgtctgtcc ggaaacaata gcggcaacgg
     5581 aaacggcagc gcgagcggcg ccggcagcgg caacaatggc aacagtagct gcaacaacag
     5641 tgccgccggg gggggcagca gcagcaacaa cacgattacc actgtagcaa attcgaatct
     5701 tgttggttct agtggctctg tctcaaattc ctctggcgtt actgctaact ctagtactgt
     5761 ttctgtagtt agttgtacag cgagtgggaa ttccataaat ggggcaggta ctgcaaacag
     5821 ttctggttca aagagtagtg caaacaattt agctagcggc cagtctagcg cttctaactt
     5881 aactaatagc accaattcaa catggcgaca aactagccaa aaccaagctc ttcaaagtca
     5941 aagcaggcca tcaggcagag aagctgactt tgattatata tctctcgttt attccattgt
     6001 tgatgattaa tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg
     6061 gtctcgattc tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     6121 cagcctcgac tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag
     6181 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     6241 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     6301 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     6361 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga
     6421 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     6481 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     6541 gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg
     6601 tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc
     6661 accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc
     6721 tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc
     6781 gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac
     6841 gaccatcagg gacagcttca aggatcgctc gcggctctta ccagccagca aaaggccagg
     6901 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6961 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7021 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7081 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     7141 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7201 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7261 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7321 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7381 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7441 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7501 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7561 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7621 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7681 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7741 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7801 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7861 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7921 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7981 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     8041 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8101 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8161 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8221 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8281 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     8341 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8401 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8461 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8521 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8581 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8641 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     8701 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgt
//
