LOCUS       sequence-290857-        7193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of 5BoxB-polyA-HhR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7193)
  AUTHORS   Kuzuoglu-Ozturk D, Bhandari D, Huntzinger E, Fauser M, Helms S, 
            Izaurralde E
  TITLE     miRISC and the CCR4-NOT complex silence mRNA targets independently 
            of 43S ribosomal scanning.
  JOURNAL   EMBO J. 2016 Jun 1;35(11):1186-203. doi: 10.15252/embj.201592901. 
            Epub 2016 Mar 23.
  PUBMED    27009120
REFERENCE   2  (bases 1 to 7193)
  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: 
            "10.15252/embj.201592901"; journalName: "EMBO J"; date: "2016-06-1- 
            1"; volume: "35"; issue: "11"; pages: "1186-203"
COMMENT     Sequence Label: pAc5.1C-no5UTR-Fluc-MtoL-V5-5BoxB-polyA-HhR_AC
FEATURES             Location/Qualifiers
     source          1..7193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2487..4139
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAEDAKNIKKGPAPFYPLEDGTAGEQLHKALKRYALVPGTIAFTD
                     AHIEVNITYAEYFELSVRLAEALKRYGLNTNHRIVVCSENSLQFFLPVLGALFIGVAVA
                     PANDIYNERELLNSLNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIILDSKTDYQGFQ
                     SLYTFVTSHLPPGFNEYDFVPESFDRDKTIALILNSSGSTGLPKGVALPHRTACVRFSH
                     ARDPIFGNQIIPDTAILSVVPFHHGFGLFTTLGYLICGFRVVLLYRFEEELFLRSLQDY
                     KIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQG
                     YGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPLIL
                     SGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESI
                     LLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTLTEKEIVDYVASQVTTAKKLRGGV
                     VFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2556..2574)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             4140..4181
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    4231..4249
                     /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    4273..4291
                     /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    4297..4315
                     /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    4355..4373
                     /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    4379..4397
                     /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             4433..4474
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4484..4501
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4524..4541)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(4598..4617)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4652..4671
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_RNA        4803..4854
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     rep_origin      complement(5375..5963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5455..5474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6134..6925,6926..6994))
                     /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     6757..6776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6995..7099)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7167..7185
                     /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 atcactatgg ccgaagacgc caaaaacata aagaaaggcc
     2521 cggcaccatt ctatccacta gaggatggaa ccgctggaga gcaactgcat aaggctttga
     2581 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtgaaca
     2641 tcacgtacgc ggaatacttc gaattgtccg ttcggttggc agaagctttg aaacgatatg
     2701 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttttgc
     2761 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
     2821 aacgtgaatt gctcaacagt ttgaacattt cgcagcctac cgtagtgttt gtttccaaaa
     2881 aggggttgca aaaaattttg aacgtgcaaa aaaaattacc aataatccag aaaattatta
     2941 tcttggattc taaaacggat taccagggat ttcagtcgtt gtacacgttc gtcacatctc
     3001 atctacctcc cggttttaat gaatacgatt ttgtaccaga gtcctttgat cgtgacaaaa
     3061 caattgcact gatattgaac tcctctggat ctactgggtt acctaagggt gtggcccttc
     3121 cgcatagaac tgcctgcgtc agattctcgc atgccagaga tcctattttt ggcaatcaaa
     3181 tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggattgttta
     3241 ctacactcgg atatttgata tgtggatttc gagtcgtctt attgtataga tttgaagaag
     3301 agctgttttt acgatccctt caggattaca aaattcaaag tgcgttgcta gtaccaaccc
     3361 tattttcatt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
     3421 aaattgcttc tgggggcgca cctctttcga aagaagtcgg ggaagcggtt gcaaaacgct
     3481 tccatcttcc agggatacga caaggatatg ggctcactga gactacatca gctattctga
     3541 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
     3601 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcagaga ggcgaattat
     3661 gtgtcagagg acctttgatt ttgtccggtt atgtaaacaa tccggaagcg accaacgcct
     3721 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac
     3781 acttcttcat agttgaccgc ttgaagtctt taattaaata caaagggtat caggtggccc
     3841 ccgctgaatt ggaatcgata ttgttacaac accccaacat cttcgacgcg ggcgtggcag
     3901 gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
     3961 agacgttgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
     4021 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
     4081 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag tccaaattgg
     4141 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gtaagaattc gctagcttcc
     4201 ctaagtccaa ctaccaaact ggggattcct gggccctgaa gaagggcccc tcgactagtc
     4261 caaatacaaa ctgggccctg aagaagggcc catatagggc cctgaagaag ggccctatcg
     4321 aggatattat ctcgactaag tccaactaca aactgggccc tgaagaaggg cccatatagg
     4381 gccctgaaga agggccctat cgaggatatt atctcgaggt cacccattcg aaggtaagcc
     4441 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     4501 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     4561 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     4621 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctgcggccgc
     4681 atacaaagga tctggctgct aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4741 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaccc
     4801 cccctgtcac cggatgtgtt ttccggtctg atgagtccgt gaggacgaaa cagggaattc
     4861 gatatcaagc ttatcgatac cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     4921 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     4981 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     5041 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     5101 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     5161 ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga
     5221 tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg
     5281 ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaaggc cagcaaaagg
     5341 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5401 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5461 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5521 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     5581 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5641 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5701 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5761 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5821 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5881 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5941 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6001 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6061 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6121 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6181 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6241 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6301 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6361 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6421 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     6481 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6541 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6601 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6661 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6721 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     6781 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6841 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6901 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6961 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7021 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7081 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7141 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgt
//
