LOCUS       sequence-356590-        7929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of 
            CeAIN2_254-400-HsTNRC6C_1207-1690-CIM1-2mut-F1389A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7929)
  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 7929)
  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-HsTNRC6C_1207-1690-CIM1-2mut-F1389A_R
FEATURES             Location/Qualifiers
     source          1..7929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /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(2630..2651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2891..2910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3238..3259
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5235..5276
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5286..5303
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5326..5343)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5376..5510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5400..5419)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5454..5473
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6111..6699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6191..6210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6870..7661,7662..7730))
                     /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     7493..7512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7731..7835)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7903..7921
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtaa atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 catcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2641 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2701 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2761 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2821 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2881 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2941 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3001 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3061 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3121 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3181 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3241 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3301 gccaagctta atggatgatt attcaatgtg gactgatgag aatgatgaag acgcgaaaaa
     3361 gaaaaaatcg ttgagagata agggattgag tggatggggt gatgctgatg tatcaaacag
     3421 taaaccaatt cgccgttgga ttgtgccaga aggccaagaa gaagactttg aaacggcaat
     3481 ggaacgatgt ccaagtcatt tgaaaaagaa agcattgaat gaagaagctc ttcgtcgacg
     3541 tttaacatct gaaaatccac aaatagtggc acaagctcaa caacaagccg aagaagaggc
     3601 gaatgctatg atgaggattg gacgaagacc cattgttccc agtggatggg gtgatattcc
     3661 atctgaaatt agtactgaca agagtgaatt cgatcaaagc tcgtcacgtg gttgggatga
     3721 aggaagtata agcggtggtg gaagttgcgg ccgcgcccag gccctgctcg tgaagcagcc
     3781 accaccgcca ccgcccccgc cgcacctgtc tctgcacccc tctgcaggca aatcggccat
     3841 ggacagcttc ccctcgcacc cacagactcc cggcctacct gacctgcaga ccaaagagca
     3901 gcagtcttca cccaacacct ttgctcctta ccctctcgct ggactgaacc caaacatgaa
     3961 tgtcaacagc atggacatga ccggtggctt gtcggtgaag gacccatccc agtccgcggc
     4021 agccgccccc gcggcgacgc accccaactc catggataac ttgcccagtg ccgcttcccc
     4081 cctggagcag aaccctagca agcatggtgc tatccctgga ggtctaagca ttgggcctcc
     4141 aggtaagtcc tccattgatg actcctatgg ccggtacgat ttaatccaga acagtgagtc
     4201 accagccagt cctcccgtag ctgttcccca tagctggtca cgtgccaaat ctgacagtga
     4261 taaaatctca aatggctcta gcatcaactg gcccccagaa gcccatccgg gagttccatg
     4321 gaaaggactg cagaatattg accctgagaa tgaccctgac gtcactcctg gcagtgtccc
     4381 cactgggcct accatcaaca ccaccatcca ggatgtcaac cgctacctcc tcaagagtgg
     4441 aggtaaactg tcagacatca aatcgacgtg gtcctctggc cctacctccc acacgcaagc
     4501 ctctctgtct catgaactat ggaaggtgcc cagaaacagt actgcaccca cgaggccacc
     4561 tccagggtta accaatccca agccctcctc cacctggggt gccagccccc tcggctggac
     4621 cagctcctac tcctcgggtt ctgcctggag caccgacacc tcaggaagaa ccagcagctg
     4681 gctcgttctt cgaaacctca ctccccagat tgatggttct acactgcgga cattgtgttt
     4741 gcaacatggg cctcttatca cattccacct gaatctgact caaggcaatg ctgtggtccg
     4801 gtacagctcc aaggaggagg ctgccaaggc ccagaagtct ctgcacatgt gcgtcctggg
     4861 aaacactacc atcctggccg agttcgctgg tgaagaagaa gtgaatcgct tcttagccca
     4921 aggccaggcg ctgccaccca cttccagctg gcagtccagc agcgcgtcca gccagccgcg
     4981 gctcagcgca gcgggcagct cccatggcct ggtacgcagc gacgctggcc actggaacgc
     5041 cccgtgcctg ggtggcaagg ggagcagtga gctggcggcg gccggggtgc cccagtactc
     5101 cagcagcgcg gcggccccgc ccagcgccga cgacagcagg gtgataggca gccccacgcc
     5161 gctaaccacc ctgctgcctg gggacctgct cagcggggag tccctgtagt aatctagagg
     5221 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     5281 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     5341 ctaaggcctg agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg
     5401 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5461 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     5521 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     5581 aaaacctcta caaatgtggt atggctgatt atgatcagtc gaccgatgcc cttgagagcc
     5641 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     5701 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     5761 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     5821 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     5881 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     5941 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     6001 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     6061 caaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6121 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6181 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6241 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6301 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6361 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6421 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6481 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6541 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6601 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6661 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6721 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6781 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6841 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6901 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6961 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7021 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7081 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7141 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg
     7201 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7261 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7321 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7381 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7441 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     7501 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7561 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7621 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7681 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7741 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7801 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7861 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7921 ccctttcgt
//
