LOCUS       sequence-289678-        8205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of CeAIN2-3xWmut.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8205)
  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 8205)
  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-CeAIN2-3xWmut_P
FEATURES             Location/Qualifiers
     source          1..8205
                     /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             5114..5125
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5490..5531
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5541..5558
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5581..5598)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5631..5765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5655..5674)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5709..5728
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6387..6975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6467..6486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7146..7937,7938..8006))
                     /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     7769..7788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8007..8111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8179..8197
                     /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 cagatatcca gcacagtggc ggccgcatga acggggacgg
     3361 ttggcaagga cctgggcgtg gaggaccaca tcacatgaat ggtggcggag gaggacgacg
     3421 tggtggaggt ggtggtatgc acaatcaaag aatacaacag aatccccatc atggtggaat
     3481 gaatcgtatg atgccacacg gattccgacc acaacaagga caatttatgc ctcaatatcc
     3541 tccaccacca ccaatgggtg gaatgcctga tgtgatggat attaatatgc gaatggatgg
     3601 tatgtcatta ggatcgacac catcaggaat tcctggacaa tatggtgata tgggtccacc
     3661 gccagtccaa caatggaatc aacctgctcc atttgtacct ccacctcaac aagacgatct
     3721 ctatgatcct tcgggatatg gtggtggagc tggtggtggt ggaggacatc atccatctca
     3781 tatgccacca tatggacaac gtggaaataa tcagaatcaa tacgggaatc gaatgcaagg
     3841 aggtcaaggt ggttatcgag ggaatcaaca gaggcagcag cagcaacagc ctcaacaggg
     3901 attcaatatg ccaccaatga tggttccacc tggtcagcct ccattttatg gtggaaatca
     3961 accgaatcat catcatcaac aaccctcatt tgctccgtcg ccattctcaa tgggtggagg
     4021 tggtggagga tcacaaggag gaccacacga acagcacgga ccaggtggaa atcagaatgg
     4081 aggattcgga gtttcatcat catcaatgga tgattattca atgtggactg atgagaatga
     4141 tgaagacgcg aaaaagaaaa aatcgttgag agataaggga ttgagtggag cgggtgatgc
     4201 tgatgtatca aacagtaaac caattcgccg ttggattgtg ccagaaggcc aagaagaaga
     4261 ctttgaaacg gcaatggaac gatgtccaag tcatttgaaa aagaaagcat tgaatgaaga
     4321 agctcttcgt cgacgtttaa catctgaaaa tccacaaata gtggcacaag ctcaacaaca
     4381 agccgaagaa gaggcgaatg ctatgatgag gattggacga agacccattg ttcccagtgg
     4441 agcgggtgat attccatctg aaattagtac tgacaagtca gaattcgatc aaagctcgtc
     4501 acgtggtgcg gatgaaggaa gtataagcgg tggtggaagt ggacgtcatc catctcatca
     4561 tcaatcaaat gataattcaa tgtggaatag tccatcagtt ggaggaggtc aatcaaatag
     4621 tgaaacacgt aatccattct ttgctcaaca catgcaacag caacaacatc aacaacaaca
     4681 tcagccacac tatggaggtt atggtgataa tggggtgctt gatggtggat cattgtggtc
     4741 tgctgctgtc atggatggat cctcaatggt accaccacat ccaacaatgg gtgtcggatc
     4801 atcagcttca tcgtcatcaa ttccaattgg catgttgatg atgagcggca gtggaaatgt
     4861 tggaggcgga ccaccatcct catcttcatt gttggacgag gatggtagcc agaagaagct
     4921 ggctgaaaat ctgaaattag ccgttcagaa gggtcatttg gatatttccc tactctcact
     4981 tccgcacatt ccgcccaacg tgctcgactt gctcaccgaa attttggcgg tcattccgcg
     5041 tttggacgac tatgaggatg agctcaaaaa attgggagaa aatcgccctt cgaactatga
     5101 cgagggagcc agcaaagagg agaaagaatc gtggatgagc aaggatcaaa agaacgatca
     5161 cgacaaagca gtaatcggag tagtgactgc aaagatcgag gtgacacaat tatctaaaaa
     5221 gatcaccgaa gctttgatcg aagctgggct tcttccacca caaccacaaa catctacatc
     5281 tgcaaataca tctaccaatc atcttaatca tagaggagga ccaccgccag gatccgatgg
     5341 aggagctggt ccaagtactt cacgaagtgg tggaggtggt gctgctggtg gagactcttt
     5401 taatagtagt ggaggtggtg gaccacctga tcattattat gattattcgt tccttggtta
     5461 gctcgagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc
     5521 tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     5581 cctcgactgt gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca
     5641 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     5701 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     5761 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     5821 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgaccg
     5881 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     5941 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     6001 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
     6061 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
     6121 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
     6181 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     6241 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     6301 catcagggac agcttcaagg atcgctcgcg gctcttacca gccagcaaaa ggccaggaac
     6361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     6601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7021 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7081 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7141 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7201 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7261 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7321 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7381 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7441 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7501 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7561 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7621 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7681 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7741 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     7801 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7861 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7921 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7981 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8101 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8161 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgt
//
