LOCUS       sequence-289997-        7539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of HsTNRC6C_1207-1609-M2WA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7539)
  AUTHORS   Zekri L, Kuzuoglu-Ozturk D, Izaurralde E
  TITLE     GW182 proteins cause PABP dissociation from silenced miRNA targets 
            in the absence of deadenylation.
  JOURNAL   EMBO J. 2013 Apr 3;32(7):1052-65. doi: 10.1038/emboj.2013.44. Epub 
            2013 Mar 5.
  PUBMED    23463101
REFERENCE   2  (bases 1 to 7539)
  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: 
            "10.1038/emboj.2013.44"; journalName: "EMBO J"; date: "2013-04-3- 
            3"; volume: "32"; issue: "7"; pages: "1052-65"
COMMENT     Sequence Label: pAc5.1B-EGFP-HsTNRC6C_1207-1609-M2WA_T
FEATURES             Location/Qualifiers
     source          1..7539
                     /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             4824..4865
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4875..4892
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4915..4932)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4965..5099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4989..5008)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5043..5062
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5721..6309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5801..5820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6480..7271,7272..7340))
                     /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     7103..7122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7341..7445)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7513..7531
                     /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 ggccgcgccc aggccctgct
     3361 cgtgaagcag ccaccaccgc caccgccccc gccgcacctg tctctgcacc cctctgcagg
     3421 caaatcggcc atggacagct tcccctcgca cccacagact cccggcctac ctgacctgca
     3481 gaccaaagag cagcagtctt cacccaacac ctttgctcct taccctctcg ctggactgaa
     3541 cccaaacatg aatgtcaaca gcatggacat gaccggtggc ttgtcggtga aggacccatc
     3601 ccagtcccag tcacgcctcc cccagtggac gcaccccaac tccatggata acttgcccag
     3661 tgccgcttcc cccctggagc agaaccctag caagcatggt gctatccctg gaggtctaag
     3721 cattgggcct ccaggtaagt cctccattga tgactcctat ggccggtacg atttaatcca
     3781 gaacagtgag tcaccagcca gtcctcccgt agctgttccc catagctggt cacgtgccaa
     3841 atctgacagt gataaaatct caaatggctc tagcatcaac tggcccccag aattccatcc
     3901 gggagttcca tggaaaggac tgcagaatat tgaccctgag aatgaccctg acgtcactcc
     3961 tggcagtgtc cccactgggc ctaccatcaa caccaccatc caggatgtca accgctacct
     4021 cctcaagagt ggaggtaaac tgtcagacat caaatcgacg gcgtcctctg gccctacctc
     4081 ccacacgcaa gcctctctgt ctcatgaact agcgaaggtg cccagaaaca gtactgcacc
     4141 cacgaggcca cctccagggt taaccaatcc caagccctcc tccaccgcgg gtgccagccc
     4201 cctcggcgcg accagctcct actcctcggg ttctgccgcg agcaccgaca cctcaggaag
     4261 aaccagcagc tggctcgttc ttcgaaacct cactccccag attgatggtt ctacactgcg
     4321 gacattgtgt ttgcaacatg ggcctcttat cacattccac ctgaatctga ctcaaggcaa
     4381 tgctgtggtc cggtacagct ccaaggagga ggctgccaag gcccagaagt ctctgcacat
     4441 gtgcgtcctg ggaaacacta ccatcctggc cgagttcgct ggtgaagaag aagtgaatcg
     4501 cttcttagcc caaggccagg cgctgccacc cacttccagc tggcagtcca gcagcgcgtc
     4561 cagccagccg cggctcagcg cagcgggcag ctcccatggc ctggtacgca gcgacgctgg
     4621 ccactggaac gccccgtgcc tgggtggcaa ggggagcagt gagctgctgt ggggcggggt
     4681 gccccagtac tccagcagcc tgtggggccc gcccagcgcc gacgacagca gggtgatagg
     4741 cagccccacg ccgctaacca ccctgctgcc tggggacctg ctcagcgggg agtccctgta
     4801 gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     4861 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     4921 ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa
     4981 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     5041 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     5101 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     5161 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg accgatgccc
     5221 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     5281 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     5341 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     5401 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     5461 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
     5521 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     5581 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     5641 ggacagcttc aaggatcgct cgcggctctt accagccagc aaaaggccag gaaccgtaaa
     5701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5881 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     5941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6121 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6181 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6421 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6781 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7081 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7441 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7501 ttaacctata aaaataggcg tatcacgagg ccctttcgt
//
