LOCUS       sequence-288687-        7546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of 5BoxB-7-Fluc-CG3548-3UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7546)
  AUTHORS   Eulalio A, Huntzinger E, Nishihara T, Rehwinkel J, Fauser M, 
            Izaurralde E
  TITLE     Deadenylation is a widespread effect of miRNA regulation.
  JOURNAL   RNA. 2009 Jan . 15(1):21-32.
  PUBMED    19029310
REFERENCE   2  (bases 1 to 7546)
  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"; journalName: "RNA. 2009 
            Jan . 15(1):21-32."
COMMENT     Sequence Label: pAc5.1C-5BoxB-7-FLuc-STOP-CG3548_E
FEATURES             Location/Qualifiers
     source          1..7546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2490..2508
                     /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    2532..2550
                     /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    2556..2574
                     /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    2614..2632
                     /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    2638..2656
                     /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             2795..4450
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVA
                     PANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQ
                     SMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSH
                     ARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDY
                     KIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQG
                     YGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIM
                     SGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESI
                     LLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGV
                     VFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2864..2882)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     rep_origin      complement(5728..6316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5808..5827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6487..7278,7279..7347))
                     /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     7110..7129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7348..7452)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7520..7538
                     /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 atcaagcttg ggccctgaag aagggcccct cgactagtcc
     2521 aaatacaaac tgggccctga agaagggccc atatagggcc ctgaagaagg gccctatcga
     2581 ggatattatc tcgactaagt ccaactacaa actgggccct gaagaagggc ccatataggg
     2641 ccctgaagaa gggccctatc gaggatatta tctcgaggtc acccattcga agcttcgttt
     2701 gagttcttgt gctgtgtgga tactcctccc gacacaaagc cgctccatca gccagcagtc
     2761 gtctaatcca gagaccccgg atcggggtac caacatggcc gaagacgcca aaaacataaa
     2821 gaaaggcccg gcaccattct atccactaga ggatggaacc gctggagagc aactgcataa
     2881 ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga
     2941 ggtgaacatc acgtacgcgg aatacttcga aatgtccgtt cggttggcag aagctatgaa
     3001 acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt
     3061 ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat
     3121 ttataatgaa cgtgaattgc tcaacagtat gaacatttcg cagcctaccg tagtgtttgt
     3181 ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aaattaccaa taatccagaa
     3241 aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt
     3301 cacatctcat ctacctcccg gttttaatga atacgatttt gtaccagagt cctttgatcg
     3361 tgacaaaaca attgcactga taatgaactc ctctggatct actgggttac ctaagggtgt
     3421 ggcccttccg catagaactg cctgcgtcag attctcgcat gccagagatc ctatttttgg
     3481 caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg
     3541 aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt
     3601 tgaagaagag ctgtttttac gatcccttca ggattacaaa attcaaagtg cgttgctagt
     3661 accaacccta ttttcattct tcgccaaaag cactctgatt gacaaatacg atttatctaa
     3721 tttacacgaa attgcttctg ggggcgcacc tctttcgaaa gaagtcgggg aagcggttgc
     3781 aaaacgcttc catcttccag ggatacgaca aggatatggg ctcactgaga ctacatcagc
     3841 tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt
     3901 ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcagagagg
     3961 cgaattatgt gtcagaggac ctatgattat gtccggttat gtaaacaatc cggaagcgac
     4021 caacgccttg attgacaagg atggatggct acattctgga gacatagctt actgggacga
     4081 agacgaacac ttcttcatag ttgaccgctt gaagtcttta attaaataca aagggtatca
     4141 ggtggccccc gctgaattgg aatcgatatt gttacaacac cccaacatct tcgacgcggg
     4201 cgtggcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga
     4261 gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac
     4321 cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg
     4381 aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagtc
     4441 caaattgtaa gaattcgggc gtggctgttg gcccatatgc atatacaata cgaagagagc
     4501 cagcaattag ggtgtaaggc atttagagga aacatcaaca gtactttaaa atttagcttg
     4561 aaatatgttt agcgaaatgt agctcttccg gtcagccaga gtaaccggaa acggaaacgt
     4621 aaacgccgga cgcgcataca atactcatcg acacactcaa gttgccactc ggacgaatgg
     4681 actgttgttt attgtgcaat tcgcacgaga cgaactcaca ggcattgcat tttcaagaaa
     4741 ttagtattac ataattgtat ccgtatctgt taagtactcc gggcgaagag atcaattcaa
     4801 aatgcggttg tgtcgtcgaa aaaaaaacaa actcgaaaat actcatgcac agactctgaa
     4861 taagttcccc tttgcagaac gcaatttact catcagtcag tattctctat ggccactgta
     4921 ttatttttat accttcattt gttagcttta agtttttttt acacaactac tacttatact
     4981 tgtgctgcca ccagttgggc tgccgtagtt gcagatcaat gtatccattg tccatacatt
     5041 gcgagaatct tttataattc gtctaagcat aagttactgt acgtcgtgtt gtatagcgct
     5101 gtaatatcct gtcacctgct tctgcttaat taatacccta tatcgggcga gagcaaggta
     5161 cgattaaacc attagataat gaaatacaaa tacatagtgt ataaacagat gaagtcgacc
     5221 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     5281 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     5341 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
     5401 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
     5461 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta
     5521 cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     5581 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     5641 ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agccagcaaa aggccaggaa
     5701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6781 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7081 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     7141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7381 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7441 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7501 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgt
//
