LOCUS       sequence-290833-        7279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of A93-5BoxB-HhR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7279)
  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 7279)
  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: pAc5.1C-Fluc-A93-5BoxB-HhR_AC
FEATURES             Location/Qualifiers
     source          1..7279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2588..4243
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVA
                     PANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQ
                     SMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSH
                     ARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDY
                     KIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQG
                     YGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIM
                     SGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESI
                     LLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGV
                     VFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2657..2675)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    4397..4415
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    4439..4457
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    4463..4481
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    4521..4539
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    4545..4563
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     CDS             4599..4640
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4650..4667
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4690..4707)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(4764..4783)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4818..4837
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_RNA        4869..4920
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     rep_origin      complement(5462..6050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5542..5561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6221..7012,7013..7081))
                     /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     6844..6863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7082..7186)
                     /gene="bla"
                     /label=AmpR promoter
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 gccgaagacg ccaaaaacat aaagaaaggc ccggcaccat tctatccact
     2641 agaggatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     2701 tggaacaatt gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt
     2761 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     2821 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     2881 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     2941 tatgaacatt tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt
     3001 gaacgtgcaa aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga
     3061 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     3121 tgaatacgat tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa
     3181 ctcctctgga tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt
     3241 cagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     3301 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     3361 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct
     3421 tcaggattac aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa
     3481 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc
     3541 acctctttcg aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg
     3601 acaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     3661 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     3721 taccgggaaa acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat
     3781 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     3841 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg
     3901 cttgaagtct ttaattaaat acaaagggta tcaggtggcc cccgctgaat tggaatcgat
     3961 attgttacaa caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc
     4021 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     4081 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     4141 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     4201 gatcctcata aaggccaaga agggcggaaa gtccaaattg taagaattcg ctagcttccc
     4261 ctaggaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4321 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaagg ccggccctaa gtccaactac
     4381 caaactgggg attcctgggc cctgaagaag ggcccctcga ctagtccaaa tacaaactgg
     4441 gccctgaaga agggcccata tagggccctg aagaagggcc ctatcgagga tattatctcg
     4501 actaagtcca actacaaact gggccctgaa gaagggccca tatagggccc tgaagaaggg
     4561 ccctatcgag gatattatct cgaggtcacc cattcgaagg taagcctatc cctaaccctc
     4621 tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc
     4681 gctgatcagc ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg
     4741 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     4801 aaaaatgctt tatttgtgaa atttgtgatg ctattgctgc ggccgcatac aaaggatctg
     4861 gctgctaacc tgtcaccgga tgtgttttcc ggtctgatga gtccgtgagg acgaaacagg
     4921 gaattcgata tcaagcttat cgataccgtc gaccgatgcc cttgagagcc ttcaacccag
     4981 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     5041 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     5101 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     5161 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     5221 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     5281 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     5341 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     5401 tcgcggctct taccagccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5461 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5521 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5581 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5641 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5701 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5761 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5821 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5881 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5941 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6001 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6061 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6301 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6361 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6421 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6481 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     6541 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6601 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6661 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6721 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6781 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     6841 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6901 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6961 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7021 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7081 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7141 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7201 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7261 gtatcacgag ccctttcgt
//
