LOCUS       sequence-289679-        7770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of CeAIN2-del254-400.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7770)
  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 7770)
  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-del254-400_P
FEATURES             Location/Qualifiers
     source          1..7770
                     /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             4679..4690
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5055..5096
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5106..5123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5146..5163)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5196..5330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5220..5239)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5274..5293
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5952..6540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6032..6051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6711..7502,7503..7571))
                     /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     7334..7353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7572..7676)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7744..7762
                     /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 catcaggacg tcatccatct catcatcaat caaatgataa
     4141 ttcaatgtgg aatagtccat cagttggagg aggtcaatca aatagtgaaa cacgtaatcc
     4201 attctttgct caacacatgc aacagcaaca acatcaacaa caacatcagc cacactatgg
     4261 aggttatggt gataatgggg tgcttgatgg tggatcattg tggtctgctg ctgtcatgga
     4321 tggatcctca atggtaccac cacatccaac aatgggtgtc ggatcatcag cttcatcgtc
     4381 atcaattcca attggcatgt tgatgatgag cggcagtgga aatgttggag gcggaccacc
     4441 atcctcatct tcattgttgg acgaggatgg tagccagaag aagctggctg aaaatctgaa
     4501 attagccgtt cagaagggtc atttggatat ttccctactc tcacttccgc acattccgcc
     4561 caacgtgctc gacttgctca ccgaaatttt ggcggtcatt ccgcgtttgg acgactatga
     4621 ggatgagctc aaaaaattgg gagaaaatcg cccttcgaac tatgacgagg gagccagcaa
     4681 agaggagaaa gaatcgtgga tgagcaagga tcaaaagaac gatcacgaca aagcagtaat
     4741 cggagtagtg actgcaaaga tcgaggtgac acaattatct aaaaagatca ccgaagcttt
     4801 gatcgaagct gggcttcttc caccacaacc acaaacatct acatctgcaa atacatctac
     4861 caatcatctt aatcatagag gaggaccacc gccaggatcc gatggaggag ctggtccaag
     4921 tacttcacga agtggtggag gtggtgctgc tggtggagac tcttttaata gtagtggagg
     4981 tggtggacca cctgatcatt attatgatta ttcgttcctt ggttagctcg agtctagagg
     5041 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     5101 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     5161 ctaaggcctg agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg
     5221 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5281 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     5341 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     5401 aaaacctcta caaatgtggt atggctgatt atgatcagtc gaccgatgcc cttgagagcc
     5461 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     5521 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     5581 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     5641 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     5701 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     5761 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     5821 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     5881 caaggatcgc tcgcggctct taccagccag caaaaggcca ggaaccgtaa aaaggccgcg
     5941 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6001 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6061 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6121 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     6181 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6241 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6301 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6361 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6421 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6481 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6541 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6601 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6661 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6721 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6781 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6841 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6901 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6961 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7021 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7081 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7141 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7201 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7261 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7321 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7381 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7441 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7501 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7561 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7621 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7681 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7741 aaaaataggc gtatcacgag gccctttcgt
//
