LOCUS       sequence-288604-        8471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of 5BoxB-7-Fluc-par6-3UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8471)
  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 8471)
  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-par6_D
FEATURES             Location/Qualifiers
     source          1..8471
                     /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(6654..7242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6734..6753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7413..8204,8205..8273))
                     /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     8036..8055
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8274..8378)
                     /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 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 gaattcgcta gcgattaacc cggcgataca gcacctccag cagacgcagc
     4501 cagaacttaa gcaacggaaa acgcgaattc atcgctgaga agtgcagagg attttcagtt
     4561 tatcagactt aactttcctt aagatcgaat aagttcgtta aactcgagtc aattatttaa
     4621 agttatacaa gttaggccaa cttattcaca tgtacactga tcgcttgtat atcgctcact
     4681 tactaaagcg actgtcatag ctcaaccagc catgatttga aaagtcacct tataaaatct
     4741 attggtatta gcctgaaagc ctgattgatt gagtaaacaa catatcggcc atatcccatt
     4801 catttcttgt taatcctggc cattcaaatt taacgttctg caaaatttcc agtccgattg
     4861 taggtgagca atatataaaa cacgattgta tatacatgtg taatcatcat acgactttcc
     4921 tccatacatt ccacgaaaca ctcaaagtca aagaaaacac acctattcat ttgcacgttt
     4981 ttgaatgcag tctcttgttt ttatcttttt tcctggcata tatatatatg tgtatttaaa
     5041 tttgtaagat ttacgtcttg ttattattct tgcttgttgt attagactag agtgagacag
     5101 ctagaggaga gagtgaaata gagagagggc atcaaatgct gaaggcagat caatttattt
     5161 aacttaacca tttgaaacgg caataattgt aacaaaatca catatacgag cacgaataat
     5221 ttataaaaaa aaaaaacatt ccattttatg tactgtgaca agtcggaaaa gcctaacaaa
     5281 attatatttc ctgatttgta agccttgttt ttaatttgag ccgatcattt tgcgtatatc
     5341 tgctatgttt tgtttagctc aatttcttat tttttcggca ttactcatta tatctgttta
     5401 ttttaaaaac catttgtaat ttttaacttt tgttttttcg tcattgtgtc caatactcgt
     5461 ccaagttctt tgttgtcatt gtcatcactt agagttatgg agaaaggata gaaaaacgag
     5521 gagataaagt taacaaaacg ttaagataca aacgaaacac caacagcgcc ttcgccttca
     5581 agagagcaat tcgatacttt ttatatatat acaagagaat atataaacga atacaatcta
     5641 tgttatgtac gtacccctgt acacacaatt gaattgaagt ctgtgtattc gctttagtaa
     5701 aattgagaaa acgccactca cggcagatgg gatgcaatgc gatctgctcg acttgaagtt
     5761 aacaatatta atatttatgg gtcgcaacgg gcataactaa ccccatacga acgtgtaccc
     5821 tgggtagata gcaagatcta ggatagacag cagcgaatct gaaactataa ctaaatgcgt
     5881 atcgccgccc aaccactccc ctcatatacc catttcacat atacccttgc attatcgaga
     5941 aatctgaact tacatcgaac aaaagtcgcc agaatatccc gatttccaag caaacgcatg
     6001 tatatatata tatatacaaa agctaatgtg tattcccgct aagcaactga aaagaccgaa
     6061 aagcaaatca cgaaagaaaa ccttatattt tgatgtatgt attattattt tttaataaac
     6121 ctaagagaac ttgtaaccgg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     6181 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     6241 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     6301 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     6361 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     6421 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg
     6481 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     6541 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     6601 cttaccagcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6661 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6721 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6781 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6841 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6901 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6961 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7021 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7081 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7141 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7201 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7261 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7321 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7381 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7441 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7501 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7561 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7621 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7681 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     7741 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7801 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7861 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7921 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7981 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     8041 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8101 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8161 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8221 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8281 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8341 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8401 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8461 agccctttcg t
//
