LOCUS       sequence-355926-        8181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of CeAIN2-W366AW390A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8181)
  AUTHORS   Kuzuoglu-Ozturk D, Huntzinger E, Schmidt S, Izaurralde E
  TITLE     The Caenorhabditis elegans GW182 protein AIN-1 interacts with PAB-1 
            and subunits of the PAN2-PAN3 and CCR4-NOT deadenylase complexes.
  JOURNAL   Nucleic Acids Res. 2012 Jul;40(12):5651-65. doi: 10.1093/nar/gks218.
            Epub 2012 Mar 7.
  PUBMED    22402495
REFERENCE   2  (bases 1 to 8181)
  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/gks218"; journalName: "Nucleic Acids Res"; date: 
            "2012-07"; volume: "40"; issue: "12"; pages: "5651-65"
COMMENT     Sequence Label: pAc5.1B-EGFP-CeAIN2-W366AW390A_P
FEATURES             Location/Qualifiers
     source          1..8181
                     /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"
     RBS             5111..5122
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5487..5528
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5538..5555
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5578..5595)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5628..5762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5652..5671)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5706..5725
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6363..6951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6443..6462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7122..7913,7914..7982))
                     /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     7745..7764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7983..8087)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8155..8173
                     /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 cgactagtga 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 gccaagcttg aattctgcag atatccagca cagtggcggc cgcatgaacg gggacggttg
     3361 gcaaggacct gggcgtggag gaccacatca catgaatggt ggcggaggag gacgacgtgg
     3421 tggaggtggt ggtatgcaca atcaaagaat acaacagaat ccccatcatg gtggaatgaa
     3481 tcgtatgatg ccacacggat tccgaccaca acaaggacaa tttatgcctc aatatcctcc
     3541 accaccacca atgggtggaa tgcctgatgt gatggatatt aatatgcgaa tggatggtat
     3601 gtcattagga tcgacaccat caggaattcc tggacaatat ggtgatatgg gtccaccgcc
     3661 agtccaacaa tggaatcaac ctgctccatt tgtacctcca cctcaacaag acgatctcta
     3721 tgatccttcg ggatatggtg gtggagctgg tggtggtgga ggacatcatc catctcatat
     3781 gccaccatat ggacaacgtg gaaataatca gaatcaatac gggaatcgaa tgcaaggagg
     3841 tcaaggtggt tatcgaggga atcaacagag gcagcagcag caacagcctc aacagggatt
     3901 caatatgcca ccaatgatgg ttccacctgg tcagcctcca ttttatggtg gaaatcaacc
     3961 gaatcatcat catcaacaac cctcatttgc tccgtcgcca ttctcaatgg gtggaggtgg
     4021 tggaggatca caaggaggac cacacgaaca gcacggacca ggtggaaatc agaatggagg
     4081 attcggagtt tcatcatcat caatggatga ttattcaatg tggactgatg agaatgatga
     4141 agacgcgaaa aagaaaaaat cgttgagaga taagggattg agtggatggg gtgatgctga
     4201 tgtatcaaac agtaaaccaa ttcgccgttg gattgtgcca gaaggccaag aagaagactt
     4261 tgaaacggca atggaacgat gtccaagtca tttgaaaaag aaagcattga atgaagaagc
     4321 tcttcgtcga cgtttaacat ctgaaaatcc acaaatagtg gcacaagctc aacaacaagc
     4381 cgaagaagag gcgaatgcta tgatgaggat tggacgaaga cccattgttc ccagtggagc
     4441 gggtgatatt ccatctgaaa ttagtactga caagagtgaa ttcgatcaaa gctcgtcacg
     4501 tggtgcggat gaaggaagta taagcggtgg tggaagtgga cgtcatccat ctcatcatca
     4561 atcaaatgat aattcaatgt ggaatagtcc atcagttgga ggaggtcaat caaatagtga
     4621 aacacgtaat ccattctttg ctcaacacat gcaacagcaa caacatcaac aacaacatca
     4681 gccacactat ggaggttatg gtgataatgg ggtgcttgat ggtggatcat tgtggtctgc
     4741 tgctgtcatg gatggatcct caatggtacc accacatcca acaatgggtg tcggatcatc
     4801 agcttcatcg tcatcaattc caattggcat gttgatgatg agcggcagtg gaaatgttgg
     4861 aggcggacca ccatcctcat cttcattgtt ggacgaggat ggtagccaga agaagctggc
     4921 tgaaaatctg aaattagccg ttcagaaggg tcatttggat atttccctac tctcacttcc
     4981 gcacattccg cccaacgtgc tcgacttgct caccgaaatt ttggcggtca ttccgcgttt
     5041 ggacgactat gaggatgagc tcaaaaaatt gggagaaaat cgcccttcga actatgacga
     5101 gggagccagc aaagaggaga aagaatcgtg gatgagcaag gatcaaaaga acgatcacga
     5161 caaagcagta atcggagtag tgactgcaaa gatcgaggtg acacaattat ctaaaaagat
     5221 caccgaagct ttgatcgaag ctgggcttct tccaccacaa ccacaaacat ctacatctgc
     5281 aaatacatct accaatcatc ttaatcatag aggaggacca ccgccaggat ccgatggagg
     5341 agctggtcca agtacttcac gaagtggtgg aggtggtgct gctggtggag actcttttaa
     5401 tagtagtgga ggtggtggac cacctgatca ttattatgat tattcgttcc ttggttagct
     5461 cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     5521 attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
     5581 cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga
     5641 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     5701 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     5761 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     5821 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgaccgatg
     5881 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     5941 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     6001 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     6061 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     6121 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc
     6181 ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc
     6241 ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat
     6301 cagggacagc ttcaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6361 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6421 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6481 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6541 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6601 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6661 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6721 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6781 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6841 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6901 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6961 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7021 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7081 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7141 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7201 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7261 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7321 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7381 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     7441 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7501 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7561 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7621 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7681 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     7741 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7801 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7861 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7921 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7981 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8041 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8101 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     8161 cgtatcacga ggccctttcg t
//
