LOCUS       sequence-289843-        7953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of CeAIN2_254-400-HsTNRC6C_1207-1690-CIM1-2mut.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7953)
  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 7953)
  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-CIM1mut-CIM2mut_R
FEATURES             Location/Qualifiers
     source          1..7953
                     /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             5238..5279
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5289..5306
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5329..5346)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5379..5513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5403..5422)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5457..5476
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6135..6723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6215..6234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6894..7685,7686..7754))
                     /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     7517..7536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7755..7859)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7927..7945
                     /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 cggccgcgcc caggccctgc tcgtgaagca
     3781 gccaccaccg ccaccgcccc cgccgcacct gtctctgcac ccctctgcag gcaaatcggc
     3841 catggacagc ttcccctcgc acccacagac tcccggccta cctgacctgc agaccaaaga
     3901 gcagcagtct tcacccaaca cctttgctcc ttaccctctc gctggactga acccaaacat
     3961 gaatgtcaac agcatggaca tgaccggtgg cttgtcggtg aaggacccat cccagtccgc
     4021 ggcagccgcc cccgcggcga cgcaccccaa ctccatggat aacttgccca gtgccgcttc
     4081 ccccctggag cagaacccta gcaagcatgg tgctatccct ggaggtctaa gcattgggcc
     4141 tccaggtaag tcctccattg atgactccta tggccggtac gatttaatcc agaacagtga
     4201 gtcaccagcc agtcctcccg tagctgttcc ccatagctgg tcacgtgcca aatctgacag
     4261 tgataaaatc tcaaatggct ctagcatcaa ctggccccca gaattccatc cgggagttcc
     4321 atggaaagga ctgcagaata ttgaccctga gaatgaccct gacgtcactc ctggcagtgt
     4381 ccccactggg cctaccatca acaccaccat ccaggatgtc aaccgctacc tcctcaagag
     4441 tggaggtaaa ctgtcagaca tcaaatcgac gtggtcctct ggccctacct cccacacgca
     4501 agcctctctg tctcatgaac tatggaaggt gcccagaaac agtactgcac ccacgaggcc
     4561 acctccaggg ttaaccaatc ccaagccctc ctccacctgg ggtgccagcc ccctcggctg
     4621 gaccagctcc tactcctcgg gttctgcctg gagcaccgac acctcaggaa gaaccagcag
     4681 ctggctcgtt cttcgaaacc tcactcccca gattgatggt tctacactgc ggacattgtg
     4741 tttgcaacat gggcctctta tcacattcca cctgaatctg actcaaggca atgctgtggt
     4801 ccggtacagc tccaaggagg aggctgccaa ggcccagaag tctctgcaca tgtgcgtcct
     4861 gggaaacact accatcctgg ccgagttcgc tggtgaagaa gaagtgaatc gcttcttagc
     4921 ccaaggccag gcgctgccac ccacttccag ctggcagtcc agcagcgcgt ccagccagcc
     4981 gcggctcagc gcagcgggca gctcccatgg cctggtacgc agcgacgctg gccactggaa
     5041 cgccccgtgc ctgggtggca aggggagcag tgagctggcg gcggccgggg tgccccagta
     5101 ctccagcagc gcggcggccc cgcccagcgc cgacgacagc agggtgatag gcagccccac
     5161 gccgctaacc accctgctgc ctggggacct gctcagcggg gagtccctgt agtaatctag
     5221 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     5281 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     5341 cttctaaggc ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca
     5401 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5461 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5521 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     5581 agtaaaacct ctacaaatgt ggtatggctg attatgatca gtcgaccgat gcccttgaga
     5641 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     5701 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     5761 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     5821 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     5881 gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg
     5941 ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc
     6001 gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag
     6061 cttcaaggat cgctcgcggc tcttaccagc cagcaaaagg ccaggaaccg taaaaaggcc
     6121 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6181 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6241 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6301 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6361 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6421 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6481 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6541 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6601 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6661 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6721 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6781 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6841 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6901 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6961 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7021 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7081 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7141 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7201 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7261 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7321 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7381 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7441 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7501 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7561 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7621 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7681 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7741 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7801 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7861 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     7921 tataaaaata ggcgtatcac gaggcccttt cgt
//
