LOCUS       sequence-333417-        7869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of 5BoxB-7-Fluc-CG10011-3UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7869)
  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 7869)
  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-5Box-7-Fluc-STOP-CG10011-SV40_D
FEATURES             Location/Qualifiers
     source          1..7869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..43
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    2538..2556
                     /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    2580..2598
                     /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    2604..2622
                     /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    2662..2680
                     /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    2686..2704
                     /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             2843..4498
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVA
                     PANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQ
                     SMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSH
                     ARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDY
                     KIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQG
                     YGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIM
                     SGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESI
                     LLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGV
                     VFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2912..2930)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(5434..5458)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      complement(6102..6690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6182..6201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6861..7652,7653..7721))
                     /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     7484..7503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7722..7826)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
       61 ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaaatcatg aatggcatca
      121 actctgaatc aaatctttgc agatgcacct acttctcatt tccactgtca catcattttt
      181 ccagatctcg ctgcctgtta tgtggcccac aaaccaagac acgttttatg gccattaaag
      241 ctggctgatc gtcgccaaac accaaataca taatgaatat gtacacattc gagaaagaag
      301 cgatcaaaga agcgtcttcg ggcggagtag gagaatgcgg aggagaagga gaacgagctg
      361 atctagtatc tctccacaat ccaatgccaa ctgaccaact ggccatattc ggagcaattt
      421 gaagccaatt tccatcgcct ggcgatcgct ccattcttgg ctatatgttt ttcaccgtta
      481 cccggggcca ttttcaaaga ctcgtcggca agataagatt gtgtcactcg ctgtctctct
      541 tcatttgtcg aagaatgctg aggaatttcg cgatgacgtc ggcgagtatt ttgaagaatg
      601 agaataattt gtatttatac gaaaatcagt tagtggaatt ttctacaaaa acatgttatc
      661 tatagataat tttgttgcaa aatatgttga ctatgacaaa gattgtatgt atataccttt
      721 aatgtattct cattttctta tgtatttata atggcaatga tgatactgat gatattttaa
      781 gatgatgcca gaccaaaagg cttgaatttc tgcgtctttt gccgaacgca gtgcatgtgc
      841 aattgttgtt ttttggaata ttcaattttc ggactgtccg ctttgatttc agtttcttgg
      901 cttattcaaa aagcaaagta aagccaaaaa agcgagatgg caataccaaa tgcggcaaaa
      961 cggtagtgga aggaaagggg tgcggggcag cggaaggaag ggtggggcgg ggcgtggcgg
     1021 ggtctgtggc tgggcgcgac gtcaccgacg ttggagccac tcctttgacc atgtgtgcgt
     1081 gtgtgtatta ttcgtgtctc gccactcgcc ggttgttttt ttctttttat gctgcgctct
     1141 ctctagcgcc atctcgctta cgcatgctca acgcaccgca tgttgccgtt tccttttatg
     1201 cgtcattttg gctcgaaata ggcaattatt taaacaaaga ttagtcaacg aaaacgctaa
     1261 aataaataag tctacaatat ggttacttat tgccatgtgt gtgcagccaa cgatagcaac
     1321 aaaagcaaca acacaggtgg ctttccctct ttcacttttt gtttgcaagc cgcgtgcgag
     1381 caagacggca cgaccggcaa acgcaattac gctgacaaag agcagacgaa gttttggcga
     1441 aaaacatcaa ggcgcctgat acgaatgcat ttgcaataac aattgcgata tttaatattg
     1501 tttatgaagc tgtttgactt caaaacacac aaaaaaaaaa ataaaacaaa ttatttgaaa
     1561 gagaattagg aatcggacgc ttatcgttag ggtaacaaca agaaatgctt actgagtcac
     1621 agcctctgga aaactgccgc aagccagaga gagagagaaa aagagggaga gcagcttaga
     1681 ccgcatgtgc ttgtgtgtga ggcgtctctc tcttcgtctc tgttgcgcaa acgcatagac
     1741 tgcactgaaa aaatcgatta cctatttttt atgaatgaat atttgcacta ttactattca
     1801 aaactattaa gatagcaatc acattcaata gccaaatact ataccacctg agcgatgcaa
     1861 cgaaatgatc aatttgagca aaaatgctgc atatttagga cggcatcatt atagaaatgc
     1921 ttcttgctgt gtacttttct ctcgtctggc agctgtttcg ccgttattgt taaaaccggc
     1981 ttaagttagg tgtgttttct acgactagtg aatgccctac tagaagatgt gtgttgcaca
     2041 aaatgtccct ggaataacca atttgaagtg cagatagcag taaacgtaag ctaatatgaa
     2101 tattatttaa ctgtaatgtt ttaatatcgc tggacattac taataaaccc actataaaca
     2161 catgtacata tgtatgtttt ggcatacaat gagtagttgg ggaaaaaatg tgtaaaagca
     2221 ccgtgaccat cacagcataa agataaccag ctgaagtatc gaatatgagt aacccccaaa
     2281 ttgaatcaca tgccgcaact gataggaccc atggaagtac actcttcatg gcgatataca
     2341 agacacacac aagcacgaac acccagttgc ggaggaaatt ctccgtaaat gaaaacccaa
     2401 tcggcgaaca attcataccc atatatggta aaagttttga acgcgacttg agagcggaga
     2461 gcattgcggc tgataaggtt ttagcgctaa gcgggcttta taaaacgggc tgcgggacca
     2521 gttttcatat caagcttggg ccctgaagaa gggcccctcg actagtccaa atacaaactg
     2581 ggccctgaag aagggcccat atagggccct gaagaagggc cctatcgagg atattatctc
     2641 gactaagtcc aactacaaac tgggccctga agaagggccc atatagggcc ctgaagaagg
     2701 gccctatcga ggatattatc tcgaggtcac ccattcgaag cttcgtttga gttcttgtgc
     2761 tgtgtggata ctcctcccga cacaaagccg ctccatcagc cagcagtcgt ctaatccaga
     2821 gaccccggat cggggtacca acatggccga agacgccaaa aacataaaga aaggcccggc
     2881 accattctat ccactagagg atggaaccgc tggagagcaa ctgcataagg ctatgaagag
     2941 atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg tgaacatcac
     3001 gtacgcggaa tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct
     3061 gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt
     3121 gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg
     3181 tgaattgctc aacagtatga acatttcgca gcctaccgta gtgtttgttt ccaaaaaggg
     3241 gttgcaaaaa attttgaacg tgcaaaaaaa attaccaata atccagaaaa ttattatcat
     3301 ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct
     3361 acctcccggt tttaatgaat acgattttgt accagagtcc tttgatcgtg acaaaacaat
     3421 tgcactgata atgaactcct ctggatctac tgggttacct aagggtgtgg cccttccgca
     3481 tagaactgcc tgcgtcagat tctcgcatgc cagagatcct atttttggca atcaaatcat
     3541 tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac
     3601 actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct
     3661 gtttttacga tcccttcagg attacaaaat tcaaagtgcg ttgctagtac caaccctatt
     3721 ttcattcttc gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat
     3781 tgcttctggg ggcgcacctc tttcgaaaga agtcggggaa gcggttgcaa aacgcttcca
     3841 tcttccaggg atacgacaag gatatgggct cactgagact acatcagcta ttctgattac
     3901 acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa
     3961 ggttgtggat ctggataccg ggaaaacgct gggcgttaat cagagaggcg aattatgtgt
     4021 cagaggacct atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat
     4081 tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag acgaacactt
     4141 cttcatagtt gaccgcttga agtctttaat taaatacaaa gggtatcagg tggcccccgc
     4201 tgaattggaa tcgatattgt tacaacaccc caacatcttc gacgcgggcg tggcaggtct
     4261 tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac
     4321 gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt
     4381 gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc
     4441 aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagtcca aattgtaaga
     4501 attcacgcac cggattaaat attaatgtgc tccccaaaaa ctcactcctt aaggcatcaa
     4561 aacacccact tgcatactca acccttaaat cgtaacttat ttgagtttat ataccttagt
     4621 acttttcaga aagtatttgt tttttgtgat agtttcaatt ttagtaaact tttgaatatg
     4681 ttctgattta actaaatata tttaggcata ctacccatac gacgtagttt aaggcatgcg
     4741 ccaggtatta agctaaaatc ccatatatag aatacttcat tacatttttg gcatgcgaac
     4801 tgtagcagat tcttcttcaa tgtagcaaaa actaacgaaa tttaaataga tcacgacacg
     4861 aatgtatact taagagagct cagactcagc actgttctca gaattaattt taaagtattt
     4921 attaaatcac attgcgtaag tcagagcatc ggcttataat ttaactataa aatataattt
     4981 atgctggaac aggcttccca aagacaaggg atcgtgctct gaaggccata ctactccaat
     5041 gcgcatttat tggcaatcgg ctatagaaac gtaatttatt tatacgagct actttaccaa
     5101 ttaattacgc tgtttatgtc aattctttaa aaccagttcc caaagtacca aaaaccgatt
     5161 atccactccc actttgtgct tttaaaggca caagaaagat cccagagata cgatatatgt
     5221 gtattatatt gctcgcacct aaaaactcaa agcggtccgt tcctgtacct taacacttgt
     5281 cactaatttt atgtaaccct taagccggac aaaccaatca aatacacata aacaactgaa
     5341 gcattaatta aatattttct gaaactctac tcccactgca aagcagcaaa tcacagacac
     5401 aacaaacaac gcactgggct aaatccaaaa aggacccagc tttcttgtac aaagtggttc
     5461 gataattcgc tagcaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     5521 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     5581 acaaatgtgg tatggctgat tatgatcagt cgaccgatgc ccttgagagc cttcaaccca
     5641 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     5701 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     5761 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     5821 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     5881 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     5941 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     6001 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggccag
     6061 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6121 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6181 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6241 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6301 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6361 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6421 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6481 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6541 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6601 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6661 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6721 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6781 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6841 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6901 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6961 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7021 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7081 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7141 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     7201 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7261 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7321 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7381 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7441 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     7501 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7561 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7621 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7681 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7741 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7801 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7861 gaaaccatt
//
