LOCUS       sequence-336056-        9770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmUPF2-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9770)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 9770)
  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"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmUPF2-siRNAres-V5His6_E
FEATURES             Location/Qualifiers
     source          1..9770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(2155..2157,2158..2160,2161..2871)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2155..2157,2158..2160,2161..2871)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2200..2221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2461..2480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2808..2829
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6646..6687
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6697..6714
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6737..6754)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6787..6921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6811..6830)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6865..6884
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7522..8110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7602..7621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8281..9072,9073..9141))
                     /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     8904..8923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9142..9246)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9314..9332
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(9396..9770,1..2135)
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc tgtctctctt
       61 catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt tgaagaatga
      121 gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa catgttatct
      181 atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta tataccttta
      241 atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg atattttaag
      301 atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag tgcatgtgca
      361 attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca gtttcttggc
      421 ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat gcggcaaaac
      481 ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg gcgtggcggg
      541 gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca tgtgtgcgtg
      601 tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg ctgcgctctc
      661 tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt ccttttatgc
      721 gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga aaacgctaaa
      781 ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac gatagcaaca
      841 aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc gcgtgcgagc
      901 aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag ttttggcgaa
      961 aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat ttaatattgt
     1021 ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat tatttgaaag
     1081 agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta ctgagtcaca
     1141 gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag cagcttagac
     1201 cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa cgcatagact
     1261 gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat tactattcaa
     1321 aactattaag atagcaatca cattcaatag ccaaatacta taccacctga gcgatgcaac
     1381 gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta tagaaatgct
     1441 tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt aaaaccggct
     1501 taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg tgttgcacaa
     1561 aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc taatatgaat
     1621 attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca ctataaacac
     1681 atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt gtaaaagcac
     1741 cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta acccccaaat
     1801 tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg cgatatacaa
     1861 gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg aaaacccaat
     1921 cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga gagcggagag
     1981 cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct gcgggaccag
     2041 ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc gacacaaagc
     2101 cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac catcatggtg
     2161 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2221 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2281 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2341 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2401 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2461 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2521 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2581 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2641 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2701 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2761 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2821 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gccaagcttg
     2881 aattctatgc tagccaacga ttctgcaacg actgacagcg aggtgtccac accgccgtcg
     2941 tcgaggaaag atttggcaga tggtgttgcc gagggtgaca acgacaacga catcgatacc
     3001 ggcgacacgg acgcaaatgc agatgaagat gtggatgcgg ctgctgccat tgccctggag
     3061 gccgaggagc gcgaggagct gcaacagttc ataagcgagc tgcgggaaaa aattgaaagt
     3121 aagcgacagc tgcgcctgca aaactccaca atcgagctgc ccggtgagga gtacttcgcc
     3181 aaactggact ccaatctcaa aaagaacacg gcattcgtca agaaactgaa actattcacg
     3241 gccacacagc tggacggact attgcgtgag atatccgctc taaacctgag caaatacatc
     3301 tcggaaatat gcgccgcgct ggtcgaggcc aaactgaaaa tgactgatgt gccggcggtg
     3361 gtgacgctgg cctctcggct gcactgcaca tatgccgact tcgatgtcca ttttctggag
     3421 gcctggcaaa aggcactgaa catcaaggcc accgagaaga ttggcaatcc gagcaaactg
     3481 cgcgttgatc ttcgcctgtt tgcggaacta gtcagctctg gtgtgatcca aatgaagccc
     3541 ggattggccc aactgggtgt tgttctggtg catttgattg ccctggacaa ggatgaccac
     3601 agcaacttct cgatcatcct ctcattctgc cggcactgcg gcgaggagta cgccggccta
     3661 gtgccgcaaa agatgcaaca gttggccacc aaatacgggg tggaggtgcc gaaatccgac
     3721 ttcctgactg ccgacaagca gctcaatctg cgcaccatgc ttaagggtta cttcaaggcc
     3781 ctgtgcaagc atgtcctggc cgagcaggcc gaactgatga acatgaccaa gaacattcgg
     3841 cgcaccatgg agtgcaaggg tgagatatcc acggagaagc gcgagaagtg tgaactgatg
     3901 caggccggat tcgataagct gctggcctcc gctcaatcgc tgtcggaact gctgggtgaa
     3961 ccactgcccg agctggccaa ggagtctgag ggctgtaatc cgggcacagt catcgacaac
     4021 atgctggaca gtgcctcctt tggtgtgctg gatccgtggg gcgacgagga gacccgagcc
     4081 ttctacacgg atttgcccga tttgagacag tttctgccaa acttttcggc acccaaagtc
     4141 gatttggaaa cgttagagga gcccagtgaa ctaaccgaag aagcgatcga tgccaatcta
     4201 gatgccgaaa tggatctaga tgatccaccg tccaccactt cggatacgac tccggagaat
     4261 ccaatcgaag agcagccaac gacgccagtt gccgctgcgg aggatctcaa gccgcagaag
     4321 atgggtaacg ctctgatgga actgggtcgc cagcagcagc agcaaagtca gctcaaccag
     4381 aatccgatcc agatccaaaa ccagatgcgt cagcagttcg atggcttcct ggtcaaccta
     4441 ttcaactgtg tgaacaagga gctgatcgat tcggcggcca tcgagttcct gcttaacttc
     4501 aataccaagc atcaacgcaa gaagctgacg cgtacgatat tctcggtgca gcgtacgcgt
     4561 ctggatatcc tgccctatct ctcccgcttc gtggccattg tgcatatgtg caataccgat
     4621 gtggccaccg atttggccga actgctgcgc aaggagttca agtggcacat tcacaagaag
     4681 aaccagctga atatcgagac gaagcttaag attgtgcgat tcattggtga actggttaag
     4741 tttggtttgt ttaaaaagtt cgatgcctta ggatgtctga agatgctgct aagggacttt
     4801 cagcatcatc aaatcgagat ggcctgtgcc tttgtggaag tcaccggcgt ctatctgtac
     4861 aattgccggg atgcccgtct gctgatgaat gtgttcctgg accaaatgct gcgcttgaaa
     4921 acagccacgg caatggactc gcgacatgcg gcgcagattg agagcgtgta ctatttggta
     4981 aagccgccgg aatcgtccaa gcgagagccc atggttcggc cggcaatgca tgagtacatc
     5041 cgttacttga tcttcgagga gctttgcaaa cagaacgttg atcgttgtat caaaatgctg
     5101 cgccgcatcg attggcagga tccggagaca aattgttatg ccattaagtg cctcagcaag
     5161 gcgtatctgc tgcgcttcca gctcattcgc tgcctggccg atctcgtatc cggactgagc
     5221 tcctatcagc cacgagcggt taccatcgtg atcgataatg tgtttgagga tatacgcgct
     5281 ggcctcgaga ttcattcgcc acgcatggca cagcggcgca tcgcgatggc caagtatctg
     5341 ggcgagatgt acaactacaa gctggtggag tccacgaaca ttcttaatac gctgtactcc
     5401 atcatctcgc tgggcgtatc gaatgaccaa aatgtcgtct cgccactgga tccgcccgat
     5461 agcctgttcc gactgaagct tgcctgtatg ctactggaca cgtgtgcacc ctacttcacc
     5521 agccaggcca cgcgaaaaaa actggactac tttctggtct tcttccaaca ctactattgg
     5581 ttcaagaaat cccatcccgt gttcagcaaa acggagaaca cctccgattt gttccccatc
     5641 ctaatggatc acacctatcg cgactgtcta ctcaatgtgc ggcccaagct gaagatttac
     5701 aagagtctag agcaggccaa ggcggcgatt gatcatctgc aggagaagct ctatccgcag
     5761 ctaaagacta ccaataatgc tcaggatccg tcgctgggca caatcagtga aataagcgag
     5821 atagacgagg gcggcactga tgaggattct gggtcatcga atgatcagcg agagcgccaa
     5881 gtgtcgggcc aagagcagga ccagagcaac gactggacag agaacgaggc ggagccaccg
     5941 ctgccgccgc ccccgccgcc agaaaagtcc aaggaggacc tggagttcga gcaattgtac
     6001 gaaaaaatga ccacagactc gtaccaagaa cggcttaagg agccgataaa ggctacggcc
     6061 aaggatatac ctgtgccgat gatggcgcgc ctgcaaaaaa aatcgtacga tcagataaca
     6121 ggtgctcaga ccatcggccc tgggatcaac aatatgtcac agatctcaaa gtcgcttccc
     6181 aatcaggacg gaacgcctgg ctctccaggt gcagattcac cagatggcaa cggcaaatcc
     6241 ggcggcggtg taggtggtag cgatgttggt gctgctggtg ctgctggtgc ttctggtgct
     6301 acatctgtac cctttgtgct aatggtgcgc ggccacaagg gcggaaagca gcaatttaag
     6361 tcatttgtgg caccatccga ctcgcacttg gccaccaacc taaagcggca ggagcagatg
     6421 atacgcgagg agaaggagaa ggtcaagcgc cttacgctta acataactga gcgcatcgag
     6481 gaggaggact accaggagtc gttgatgccg ccgcagcatc gaagcttcac ccagagctac
     6541 taccagaagc ccaataagca caagttcaag caccaaaagg gtgcgcccga tgcggacctc
     6601 atatttcacg gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct
     6661 aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt
     6721 taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg atcagcctcg
     6781 atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     6841 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     6901 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     6961 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     7021 tatgatcagt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     7081 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     7141 acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac
     7201 gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt
     7261 cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc
     7321 cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat
     7381 tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag
     7441 gcaggtagat gacgaccatc agggacagct tcaaggccag caaaaggcca ggaaccgtaa
     7501 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7561 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7621 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7681 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7741 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7801 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7861 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7921 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7981 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8041 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8101 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8161 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8221 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8281 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8341 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8401 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8461 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8521 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8581 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8641 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8701 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8761 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8821 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8881 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     8941 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9001 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9061 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9121 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9181 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9241 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     9301 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat taattctata
     9361 ttctaaaaac acaaatgata cttctaaaaa aaaatcatga atggcatcaa ctctgaatca
     9421 aatctttgca gatgcaccta cttctcattt ccactgtcac atcatttttc cagatctcgc
     9481 tgcctgttat gtggcccaca aaccaagaca cgttttatgg ccattaaagc tggctgatcg
     9541 tcgccaaaca ccaaatacat aatgaatatg tacacattcg agaaagaagc gatcaaagaa
     9601 gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag aacgagctga tctagtatct
     9661 ctccacaatc caatgccaac tgaccaactg gccatattcg gagcaatttg aagccaattt
     9721 ccatcgcctg gcgatcgctc cattcttggc tatatgtttt tcaccgttac
//
