LOCUS       sequence-288606-        8475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of 5BoxB-73-Fluc-par6-3UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8475)
  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 8475)
  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-73-Fluc-par6_D
FEATURES             Location/Qualifiers
     source          1..8475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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    2598..2616
                     /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    2622..2640
                     /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    2680..2698
                     /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    2704..2722
                     /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             2799..4454
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVA
                     PANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQ
                     SMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSH
                     ARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDY
                     KIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQG
                     YGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIM
                     SGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESI
                     LLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGV
                     VFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2868..2886)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     rep_origin      complement(6658..7246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6738..6757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7417..8208,8209..8277))
                     /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     8040..8059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8278..8382)
                     /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 agcttgggcc ctgaagaagg gcccctcgac
     2581 tagtccaaat acaaactggg ccctgaagaa gggcccatat agggccctga agaagggccc
     2641 tatcgaggat attatctcga ctaagtccaa ctacaaactg ggccctgaag aagggcccat
     2701 atagggccct gaagaagggc cctatcgagg atattatctc gaggtcaccc attcgaagct
     2761 tgtcgtctaa tccagagacc ccggatcggg gtaccaacat ggccgaagac gccaaaaaca
     2821 taaagaaagg cccggcacca ttctatccac tagaggatgg aaccgctgga gagcaactgc
     2881 ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata
     2941 tcgaggtgaa catcacgtac gcggaatact tcgaaatgtc cgttcggttg gcagaagcta
     3001 tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc
     3061 aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg
     3121 acatttataa tgaacgtgaa ttgctcaaca gtatgaacat ttcgcagcct accgtagtgt
     3181 ttgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaaatta ccaataatcc
     3241 agaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt
     3301 tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtacca gagtcctttg
     3361 atcgtgacaa aacaattgca ctgataatga actcctctgg atctactggg ttacctaagg
     3421 gtgtggccct tccgcataga actgcctgcg tcagattctc gcatgccaga gatcctattt
     3481 ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt
     3541 ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata
     3601 gatttgaaga agagctgttt ttacgatccc ttcaggatta caaaattcaa agtgcgttgc
     3661 tagtaccaac cctattttca ttcttcgcca aaagcactct gattgacaaa tacgatttat
     3721 ctaatttaca cgaaattgct tctgggggcg cacctctttc gaaagaagtc ggggaagcgg
     3781 ttgcaaaacg cttccatctt ccagggatac gacaaggata tgggctcact gagactacat
     3841 cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc
     3901 cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaga
     3961 gaggcgaatt atgtgtcaga ggacctatga ttatgtccgg ttatgtaaac aatccggaag
     4021 cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata gcttactggg
     4081 acgaagacga acacttcttc atagttgacc gcttgaagtc tttaattaaa tacaaagggt
     4141 atcaggtggc ccccgctgaa ttggaatcga tattgttaca acaccccaac atcttcgacg
     4201 cgggcgtggc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt
     4261 tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa
     4321 caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta
     4381 ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa
     4441 agtccaaatt gtaagaattc gctagcgatt aacccggcga tacagcacct ccagcagacg
     4501 cagccagaac ttaagcaacg gaaaacgcga attcatcgct gagaagtgca gaggattttc
     4561 agtttatcag acttaacttt ccttaagatc gaataagttc gttaaactcg agtcaattat
     4621 ttaaagttat acaagttagg ccaacttatt cacatgtaca ctgatcgctt gtatatcgct
     4681 cacttactaa agcgactgtc atagctcaac cagccatgat ttgaaaagtc accttataaa
     4741 atctattggt attagcctga aagcctgatt gattgagtaa acaacatatc ggccatatcc
     4801 cattcatttc ttgttaatcc tggccattca aatttaacgt tctgcaaaat ttccagtccg
     4861 attgtaggtg agcaatatat aaaacacgat tgtatataca tgtgtaatca tcatacgact
     4921 ttcctccata cattccacga aacactcaaa gtcaaagaaa acacacctat tcatttgcac
     4981 gtttttgaat gcagtctctt gtttttatct tttttcctgg catatatata tatgtgtatt
     5041 taaatttgta agatttacgt cttgttatta ttcttgcttg ttgtattaga ctagagtgag
     5101 acagctagag gagagagtga aatagagaga gggcatcaaa tgctgaaggc agatcaattt
     5161 atttaactta accatttgaa acggcaataa ttgtaacaaa atcacatata cgagcacgaa
     5221 taatttataa aaaaaaaaaa cattccattt tatgtactgt gacaagtcgg aaaagcctaa
     5281 caaaattata tttcctgatt tgtaagcctt gtttttaatt tgagccgatc attttgcgta
     5341 tatctgctat gttttgttta gctcaatttc ttattttttc ggcattactc attatatctg
     5401 tttattttaa aaaccatttg taatttttaa cttttgtttt ttcgtcattg tgtccaatac
     5461 tcgtccaagt tctttgttgt cattgtcatc acttagagtt atggagaaag gatagaaaaa
     5521 cgaggagata aagttaacaa aacgttaaga tacaaacgaa acaccaacag cgccttcgcc
     5581 ttcaagagag caattcgata ctttttatat atatacaaga gaatatataa acgaatacaa
     5641 tctatgttat gtacgtaccc ctgtacacac aattgaattg aagtctgtgt attcgcttta
     5701 gtaaaattga gaaaacgcca ctcacggcag atgggatgca atgcgatctg ctcgacttga
     5761 agttaacaat attaatattt atgggtcgca acgggcataa ctaaccccat acgaacgtgt
     5821 accctgggta gatagcaaga tctaggatag acagcagcga atctgaaact ataactaaat
     5881 gcgtatcgcc gcccaaccac tcccctcata tacccatttc acatataccc ttgcattatc
     5941 gagaaatctg aacttacatc gaacaaaagt cgccagaata tcccgatttc caagcaaacg
     6001 catgtatata tatatatata caaaagctaa tgtgtattcc cgctaagcaa ctgaaaagac
     6061 cgaaaagcaa atcacgaaag aaaaccttat attttgatgt atgtattatt attttttaat
     6121 aaacctaaga gaacttgtaa ccggtcgacc gatgcccttg agagccttca acccagtcag
     6181 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     6241 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     6301 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     6361 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     6421 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg
     6481 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
     6541 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc
     6601 ggctcttacc agccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6661 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6721 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6781 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6841 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6901 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6961 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7021 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7081 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7141 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7201 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7261 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7321 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7381 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7441 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7501 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7561 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7621 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7681 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat
     7741 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7801 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7861 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7921 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7981 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga
     8041 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8101 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8161 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8221 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8281 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8341 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8401 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8461 cacgagccct ttcgt
//
