LOCUS       sequence-290490-        9782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmPAN2-F550EM554EK557E-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9782)
  AUTHORS   Jonas S, Christie M, Peter D, Bhandari D, Loh B, Huntzinger E, 
            Weichenrieder O, Izaurralde E
  TITLE     An asymmetric PAN3 dimer recruits a single PAN2 exonuclease to 
            mediate mRNA deadenylation and decay.
  JOURNAL   Nat Struct Mol Biol. 2014 Jul;21(7):599-608. doi: 10.1038/nsmb.2837.
            Epub 2014 Jun 1.
  PUBMED    24880343
REFERENCE   2  (bases 1 to 9782)
  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"; doi: "10.1038/nsmb.2837";
            journalName: "Nat Struct Mol Biol"; date: "2014-07"; volume: "21"; 
            issue: "7"; pages: "599-608"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmPAN2-F550EM554EK557E-dsRNAres_Y
FEATURES             Location/Qualifiers
     source          1..9782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /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(2633..2654)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2894..2913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7067..7108
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7118..7135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7158..7175)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7208..7342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7232..7251)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7286..7305
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7964..8552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8044..8063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8723..9514,9515..9583))
                     /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     9346..9365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9584..9688)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9756..9774
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
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 gtcgtctaat ccagagaccc cggatcgggg
     2581 taccaacatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3301 caagccaagc ttgaattctg cagatatcat ggactatgtc tactgtggaa cggatccgat
     3361 tggagctagc gaggatatcc tgtccgtgta cgatgccggc agtgcccccg gcaatggtca
     3421 tttcagcccg tcctttaacg gctttaatat tggaacgacg gaccccgagt acgtcgaact
     3481 ggtcccggtc ctggctgatg gaggtgaaca ttttggagtg agctccgtcg cctttgatga
     3541 ttatgaggaa ttgttgtgga tgggaaatca aggaggacat gtgaccagct attataccaa
     3601 tagcatgcaa aaatatacca gcttccaagt gcatgctacg gatatcgtcc gtgatattag
     3661 cacgttggat agcggcgtct tggccctgac ccaaacgagc ttgcgccacc aaattcgtcg
     3721 cggcttgccg aaattcacgt ttaagagcaa taatatgaaa gaaatggtct ccatgttgca
     3781 actgtcgccc caccgcttgg tgatggctgg cttgcaagat gaactgattg attttgattt
     3841 gcgtaccctg aaagagacgc gtattgaaca cgtcggcgct ggaggctgta ccgtcttgcg
     3901 taaaaactcg cgctacttgt tcgctggaga tcaattgggc accgttaccc tgcgtgattt
     3961 gaattccttg tccgtgcaac ataccattaa aacccatacg aatattctga gcgatttttc
     4021 cgtgcaaggc aacttgctga ttagttgtgg ctacagcgga aggcaaaata acctggccat
     4081 cgaccgcttc ctaatggtct acgatctgcg catgctgcgc ctgatcgcgc ccattcaagt
     4141 gatgattgat ccgcagatgt tgaagttcct gccatcgctg acatcccaac tggccgtcgt
     4201 gtccagctac ggccaggttc agctggtgga cactgtggag ctcagcgagc cgcgagtatc
     4261 catgtaccag atcaacacca acggcagcca gtgcctcagc ttcgacatta gctcctcttc
     4321 acaggccatg gcatttgggg accagtcggg ccacatcaac atgattgccg ctgtacaaac
     4381 gcctcagccg cagttcaatc tctactcgcg aagcacagag tttgcggacg tggtgccaca
     4441 gctgccgatg gtctccatta cggacaccaa ctttccgcta tcgtcggtca tgctaccgca
     4501 cttgacgacg ggcacacaat ggttctcaga ttggccggag gaactgctgc gctaccgcta
     4561 ccacagaccc aagaccattg atcccgaagt gctgagcaac atgaaaatgc agggaccgat
     4621 tggatactcg ccaaacccac gcaccgcccg tcgtaatcaa attccgtacg tgattgagca
     4681 gggcggcgtg tgcagtccga acggaaatgg aactgcggcg gcgaccaagg cggagaacgg
     4741 tgtgaaaatc attccacggc ggtatcgcaa agtcgagctg aagtacacga agctgggcac
     4801 ccaggacttt gactttgacc agcataatca aacatgtttc gcggggctgg aggcaacgct
     4861 accgaactcc tactgcaatg ctatgctaca gattttgtat ttcaccgacg cgcttcgcgt
     4921 gaagctgctg gagcattcgt gcatcaagga gttttgcctg tcctgcgaac taggggaact
     4981 cttcaacgag ctggatgaaa gtaccgcttc ttcaccttgc caggccagca atttcctgcg
     5041 atctttccgc actgtgccag aggcctcggc actgggattg attctcaccg accgctcatc
     5101 gaacgtcaat ctaatatcgc tgatacagaa ctggaaccga ttcattttgc atcaaatgca
     5161 ttacgaaata ttcgattcta gcaagaacgc atccacctac agcggatctg tgcaaacatc
     5221 aacgaacgcg gaaaacgcgg gctcttcaga aacaagtggc tcctcggacc tttacgattc
     5281 cataagtgat gagaactcca aggaggacga tcgggaacgc agtaagatca acgcagaaac
     5341 tgacatcagt aagatctttg ggaccaagca aatctgtata aaccgctgca tcaaatgcca
     5401 agaagagaag ataaaggaaa gcattctact tgcctgcaat ttgtcgtatc ccaaccacat
     5461 caaagatagt gatcagtatt tcaactttgg caccattttg aagcgatctc tcagctccga
     5521 gaagagtatt caggccttct gcgagagatg taagaagttt tcgccaacca atcagagcgt
     5581 gaaggtgact tcgcttccgc aaattctgtc cattaactgt ggcctcaaca acgaaaaaga
     5641 cattaccttc ctgaagcgcc agctcaatcg ctgcagcgaa aagacgactg ttgatgcggc
     5701 cgcttccttg agcacaagca agccgtgtcg atatggagcc aactgttcgc gaagtgactg
     5761 ccatttcatg catccggatc gaaaatctcc atcgcacaca agccaaccga atgctgtcaa
     5821 caattctccc aatggacgcc aaaagtcctg gtttcccctt acatttacta tgggcatcaa
     5881 tgaccaaggt gaggtgcagg tgcaaactca atcagatgca agcagtggca aatcggaaca
     5941 ggaggaggaa acggaaaagc caccaacaaa gggcttggac aacaatcgga tgtacgcgct
     6001 gcatgccgtg gtgtgccaag tcgatgatgg tacccaaaag aacttggtat cgttgataaa
     6061 tgtgcagcgg ccataccaca ccatgaaact ggctgaatct gcagatgacc ctcagagtca
     6121 atggtatatt tttaatgatt ttagcatatc gcccgtttcg ccacaagaat cggtatggtt
     6181 cacattggat tggaaagttc cttgcatact cttctacaga cacgtagagg atgactctga
     6241 gtctgctagt actactagct cgaccgtcac ggaaagtgaa gaaaccattc catcagagag
     6301 cagcagcggc tcgccaacaa atctatccaa cccttttcta gaggaaattg tgagtcctat
     6361 gctaggcaac ttaagtgcag acgccacgct gcagcccctg cagtcagatg aaatgccgca
     6421 gtcgggtgat ctggtcgcaa tggatgccga gttcgttacc ctaaatccag aggaaaatga
     6481 gattcgcccg gatggcaaaa cggcgaccat taagccatgc cacatgagtg tggcccggat
     6541 ttcatgtata cgaggacaag gcccggctga aggagttcca ttcatggacg attacatatc
     6601 cacccaggag aaggtcgtgg actacctaac tcagttctcg ggaatcaagc ctggcgactt
     6661 ggacgctaac ttcagcaaaa agcgactgac tgccttgaag tactcatacc agaaattaaa
     6721 atacctcgtt gacgtaggag taatatttgt cggacatggt ctgaagaacg actttagggt
     6781 cataaacatc tatgtaccgt cggagcagat aatagatacg gtgcacctgt ttcacatgcc
     6841 acatcatcgc atggtttccc taagatttct agcctggcat ttcttgggta ctaagattca
     6901 gtccgagacg cacgactcca tcgaagatgc acggaccact ctgcagctct acaagcacta
     6961 tttgaagctg caggaggaga agaagtttgc caacgctctc aagaatcttt acgaacgcgg
     7021 aaagcagctg cagtggaagg ttccagagga ctgaccgcgg ttcgaaggta agcctatccc
     7081 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     7141 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     7201 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     7261 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     7321 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     7381 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     7441 ttatgatcag tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     7501 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     7561 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga
     7621 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg
     7681 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg
     7741 ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca
     7801 ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca
     7861 ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc
     7921 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7981 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8041 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8101 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     8161 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8221 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8281 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8341 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8401 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8461 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8521 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     8581 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     8641 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     8701 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8761 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8821 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8881 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8941 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9001 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     9061 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9121 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     9181 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9241 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9301 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     9361 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     9421 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     9481 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     9541 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     9601 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     9661 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     9721 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     9781 gt
//
