LOCUS       sequence-290484-        8240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmPAN2-del468-971-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8240)
  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 8240)
  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-del468-981-dsRNAres_Y
FEATURES             Location/Qualifiers
     source          1..8240
                     /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             5525..5566
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5576..5593
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5616..5633)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5666..5800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5690..5709)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5744..5763
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6422..7010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6502..6521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7181..7972,7973..8041))
                     /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     7804..7823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8042..8146)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8214..8232
                     /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 ccacaggccc aagaccattg atcccgaagt gctgagcaac atgaaaatgc agggaccgat
     4621 tggatactcg ccaaacccac gcaccgcccg tcgtaatcaa attccgtacg tgattgagca
     4681 gggcggcgtg tgcagtccga acggaaatgg aactgcggcg gcgaccaagg aaagtgaaga
     4741 aaccattcca tcagagagca gcagcggctc gccaacaaat ctatccaacc cttttctaga
     4801 ggaaattgtg agtcctatgc taggcaactt aagtgcagac gccacgctgc agcccctgca
     4861 gtcagatgaa atgccgcagt cgggtgatct ggtcgcaatg gatgccgagt tcgttaccct
     4921 aaatccagag gaaaatgaga ttcgcccgga tggcaaaacg gcgaccatta agccatgcca
     4981 catgagtgtg gcccggattt catgtatacg aggacaaggc ccggctgaag gagttccatt
     5041 catggacgat tacatatcca cccaggagaa ggtcgtggac tacctaactc agttctcggg
     5101 aatcaagcct ggcgacttgg acgctaactt cagcaaaaag cgactgactg ccttgaagta
     5161 ctcataccag aaattaaaat acctcgttga cgtaggagta atatttgtcg gacatggtct
     5221 gaagaacgac tttagggtca taaacatcta tgtaccgtcg gagcagataa tagatacggt
     5281 gcacctgttt cacatgccac atcatcgcat ggtttcccta agatttctag cctggcattt
     5341 cttgggtact aagattcagt ccgagacgca cgactccatc gaagatgcac ggaccactct
     5401 gcagctctac aagcactatt tgaagctgca ggaggagaag aagtttgcca acgctctcaa
     5461 gaatctttac gaacgcggaa agcagctgca gtggaaggtt ccagaggact gaccgcggtt
     5521 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     5581 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     5641 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     5701 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     5761 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     5821 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     5881 caaatgtggt atggctgatt atgatcagtc gaccgatgcc cttgagagcc ttcaacccag
     5941 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     6001 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     6061 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     6121 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     6181 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     6241 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     6301 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     6361 tcgcggctct taccagccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6421 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6541 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6601 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6661 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6721 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6781 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6841 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6901 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6961 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7021 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7081 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7141 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7261 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7321 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7381 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     7501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7621 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7681 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     7801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7861 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8041 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8101 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8161 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8221 gtatcacgag gccctttcgt
//
