LOCUS       sequence-288602-        7896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of 5BoxB-73-Fluc-CG10011-3UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7896)
  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 7896)
  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-73-Fluc-STOP-CG10011-SV40_D
FEATURES             Location/Qualifiers
     source          1..7896
                     /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"
     protein_bind    complement(5390..5414)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      complement(6079..6667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6159..6178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6838..7629,7630..7698))
                     /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     7461..7480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7699..7803)
                     /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 acgcaccgga ttaaatatta atgtgctccc caaaaactca
     4501 ctccttaagg catcaaaaca cccacttgca tactcaaccc ttaaatcgta acttatttga
     4561 gtttatatac cttagtactt ttcagaaagt atttgttttt tgtgatagtt tcaattttag
     4621 taaacttttg aatatgttct gatttaacta aatatattta ggcatactac ccatacgacg
     4681 tagtttaagg catgcgccag gtattaagct aaaatcccat atatagaata cttcattaca
     4741 tttttggcat gcgaactgta gcagattctt cttcaatgta gcaaaaacta acgaaattta
     4801 aatagatcac gacacgaatg tatacttaag agagctcaga ctcagcactg ttctcagaat
     4861 taattttaaa gtatttatta aatcacattg cgtaagtcag agcatcggct tataatttaa
     4921 ctataaaata taatttatgc tggaacaggc ttcccaaaga caagggatcg tgctctgaag
     4981 gccatactac tccaatgcgc atttattggc aatcggctat agaaacgtaa tttatttata
     5041 cgagctactt taccaattaa ttacgctgtt tatgtcaatt ctttaaaacc agtttccaaa
     5101 gtaccaaaaa ccgattatcc actcccactt tgtgctttta aaggcacaag aaagatccca
     5161 gagatacgat atatgtgtat tatattgctc gcacctaaaa actcaaagcg gtccgttcct
     5221 gtaccttaac acttgtcact aattttatgt aacccttaag ccggacaaac caatcaaata
     5281 cacataaaca actgaagcat taattaaata ttttctgaaa ctctactccc actgcaaagc
     5341 agcaaatcac agacacaaca aacaacgcac tgggctaaat ccaaaaagga cccagctttc
     5401 ttgtacaaag tggttcgata attcgctagc accattataa gctgcaataa acaagttaac
     5461 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     5521 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac cgatgccctt
     5581 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     5641 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     5701 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     5761 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     5821 cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct
     5881 ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg
     5941 catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg
     6001 acagcttcaa ggatcgctcg cggctcttac cagccagcaa aaggccagga accgtaaaaa
     6061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6241 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6481 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7141 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7441 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7801 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7861 aacctataaa aataggcgta tcacgagccc tttcgt
//
