LOCUS       sequence-356070-        9758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmPAN2-K1124EY1127EK1131E-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9758)
  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 9758)
  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-K1124EY1127EK1131E-dsRNAres_Y
FEATURES             Location/Qualifiers
     source          1..9758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..2510
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2530..2532,2533..2535,2536..3246)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2530..2532,2533..2535,2536..3246)
                     /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(2575..2596)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2836..2855)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3183..3204
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7009..7050
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7060..7077
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7100..7117)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7150..7284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7174..7193)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7228..7247
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7885..8473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7965..7984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8644..9435,9436..9504))
                     /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     9267..9286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9505..9609)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9677..9695
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catgaatggc atcaactctg aatcaaatct ttgcagatgc acctacttct catttccact
       61 gtcacatcat ttttccagat ctcgctgcct gttatgtggc ccacaaacca agacacgttt
      121 tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacataatga atatgtacac
      181 attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga gtaggagaat gcggaggaga
      241 aggagaacga gctgatctag tatctctcca caatccaatg ccaactgacc aactggccat
      301 attcggagca atttgaagcc aatttccatc gcctggcgat cgctccattc ttggctatat
      361 gtttttcacc gttacccggg gccattttca aagactcgtc ggcaagataa gattgtgtca
      421 ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag
      481 tattttgaag aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac
      541 aaaaacatgt tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt
      601 atgtatatac ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac
      661 tgatgatatt ttaagatgat gccagaccaa aaggcttgaa tttctgcgtc ttttgccgaa
      721 cgcagtgcat gtgcaattgt tgttttttgg aatattcaat tttcggactg tccgctttga
      781 tttcagtttc ttggcttatt caaaaagcaa agtaaagcca aaaaagcgag atggcaatac
      841 caaatgcggc aaaacggtag tggaaggaaa ggggtgcggg gcagcggaag gaagggtggg
      901 gcggggcgtg gcggggtctg tggctgggcg cgacgtcacc gacgttggag ccactccttt
      961 gaccatgtgt gcgtgtgtgt attattcgtg tctcgccact cgccggttgt ttttttcttt
     1021 ttatgctgcg ctctctctag cgccatctcg cttacgcatg ctcaacgcac cgcatgttgc
     1081 cgtttccttt tatgcgtcat tttggctcga aataggcaat tatttaaaca aagattagtc
     1141 aacgaaaacg ctaaaataaa taagtctaca atatggttac ttattgccat gtgtgtgcag
     1201 ccaacgatag caacaaaagc aacaacacag gtggctttcc ctctttcact ttttgtttgc
     1261 aagccgcgtg cgagcaagac ggcacgaccg gcaaacgcaa ttacgctgac aaagagcaga
     1321 cgaagttttg gcgaaaaaca tcaaggcgcc tgatacgaat gcatttgcaa taacaattgc
     1381 gatatttaat attgtttatg aagctgtttg acttcaaaac acacaaaaaa aaaaataaaa
     1441 caaattattt gaaagagaat taggaatcgg acgcttatcg ttagggtaac aacaagaaat
     1501 gcttactgag tcacagcctc tggaaaactg ccgcaagcca gagagagaga gaaaaagagg
     1561 gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg tctctgttgc
     1621 gcaaacgcat agactgcact gaaaaaatcg attacctatt ttttatgaat gaatatttgc
     1681 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1741 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1801 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1861 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgaatgcc ctactagaag
     1921 atgtgtgttg cacaaaatgt ccctggaata accaatttga agtgcagata gcagtaaacg
     1981 taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca ttactaataa
     2041 acccactata aacacatgta catatgtatg ttttggcata caatgagtag ttggggaaaa
     2101 aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag tatcgaatat
     2161 gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa gtacactctt
     2221 catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga aattctccgt
     2281 aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt ttgaacgcga
     2341 cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc tttataaaac
     2401 gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt gtggatactc
     2461 ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac cccggatcgg
     2521 ggtaccatca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2581 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2641 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2701 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2761 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2821 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2881 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2941 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3001 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3061 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3121 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3181 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3241 tacaagccaa gcttgaattc tgcagatatc atggactatg tctactgtgg aacggatccg
     3301 attggagcta gcgaggatat cctgtccgtg tacgatgccg gcagtgcccc cggcaatggt
     3361 catttcagcc cgtcctttaa cggctttaat attggaacga cggaccccga gtacgtcgaa
     3421 ctggtcccgg tcctggctga tggaggtgaa cattttggag tgagctccgt cgcctttgat
     3481 gattatgagg aattgttgtg gatgggaaat caaggaggac atgtgaccag ctattatacc
     3541 aatagcatgc aaaaatatac cagcttccaa gtgcatgcta cggatatcgt ccgtgatatt
     3601 agcacgttgg atagcggcgt cttggccctg acccaaacga gcttgcgcca ccaaattcgt
     3661 cgcggcttgc cgaaattcac gtttaagagc aataatatga aagaaatggt ctccatgttg
     3721 caactgtcgc cccaccgctt ggtgatggct ggcttgcaag atgaactgat tgattttgat
     3781 ttgcgtaccc tgaaagagac gcgtattgaa cacgtcggcg ctggaggctg taccgtcttg
     3841 cgtaaaaact cgcgctactt gttcgctgga gatcaattgg gcaccgttac cctgcgtgat
     3901 ttgaattcct tgtccgtgca acataccatt aaaacccata cgaatattct gagcgatttt
     3961 tccgtgcaag gcaacttgct gattagttgt ggctacagcg gaaggcaaaa taacctggcc
     4021 atcgaccgct tcctaatggt ctacgatctg cgcatgctgc gcctgatcgc gcccattcaa
     4081 gtgatgattg atccgcagat gttgaagttc ctgccatcgc tgacatccca actggccgtc
     4141 gtgtccagct acggccaggt tcagctggtg gacactgtgg agctcagcga gccgcgagta
     4201 tccatgtacc agatcaacac caacggcagc cagtgcctca gcttcgacat tagctcctct
     4261 tcacaggcca tggcatttgg ggaccagtcg ggccacatca acatgattgc cgctgtacaa
     4321 acgcctcagc cgcagttcaa tctctactcg cgaagcacag agtttgcgga cgtggtgcca
     4381 cagctgccga tggtctccat tacggacacc aactttccgc tatcgtcggt catgctaccg
     4441 cacttgacga cgggcacaca atggttctca gattggccgg aggaactgct gcgctaccgc
     4501 taccacagac ccaagaccat tgatcccgaa gtgctgagca acatgaaaat gcagggaccg
     4561 attggatact cgccaaaccc acgcaccgcc cgtcgtaatc aaattccgta cgtgattgag
     4621 cagggcggcg tgtgcagtcc gaacggaaat ggaactgcgg cggcgaccaa ggcggagaac
     4681 ggtgtgaaaa tcattccacg gcggtatcgc aaagtcgagc tgaagtacac gaagctgggc
     4741 acccaggact ttgactttga ccagcataat caaacatgtt tcgcggggct ggaggcaacg
     4801 ctaccgaact cctactgcaa tgctatgcta cagattttgt atttcaccga cgcgcttcgc
     4861 gtgaagctgc tggagcattc gtgcatcaag gagttttgcc tgtcctgcga actagggttc
     4921 ctcttcaaca tgctggataa aagtaccgct tcttcacctt gccaggccag caatttcctg
     4981 cgatctttcc gcactgtgcc agaggcctcg gcactgggat tgattctcac cgaccgctca
     5041 tcgaacgtca atctaatatc gctgatacag aactggaacc gattcatttt gcatcaaatg
     5101 cattacgaaa tattcgattc tagcaagaac gcatccacct acagcggatc tgtgcaaaca
     5161 tcaacgaacg cggaaaacgc gggctcttca gaaacaagtg gctcctcgga cctttacgat
     5221 tccataagtg atgagaactc caaggaggac gatcgggaac gcagtaagat caacgcagaa
     5281 actgacatca gtaagatctt tgggaccaag caaatctgta taaaccgctg catcaaatgc
     5341 caagaagaga agataaagga aagcattcta cttgcctgca atttgtcgta tcccaaccac
     5401 atcaaagata gtgatcagta tttcaacttt ggcaccattt tgaagcgatc tctcagctcc
     5461 gagaagagta ttcaggcctt ctgcgagaga tgtaagaagt tttcgccaac caatcagagc
     5521 gtgaaggtga cttcgcttcc gcaaattctg tccattaact gtggcctcaa caacgaaaaa
     5581 gacattacct tcctgaagcg ccagctcaat cgctgcagcg aaaagacgac tgttgatgcg
     5641 gccgcttcct tgagcacaag caagccgtgt cgatatggag ccaactgttc gcgaagtgac
     5701 tgccatttca tgcatccgga tcgaaaatct ccatcgcaca caagccaacc gaatgctgtc
     5761 aacaattctc ccaatggacg ccaaaagtcc tggtttcccc ttacatttac tatgggcatc
     5821 aatgaccaag gtgaggtgca ggtgcaaact caatcagatg caagcagtgg caaatcggaa
     5881 caggaggagg aaacggaaaa gccaccaaca aagggcttgg acaacaatcg gatgtacgcg
     5941 ctgcatgccg tggtgtgcca agtcgatgat ggtacccaaa agaacttggt atcgttgata
     6001 aatgtgcagc ggccatacca caccatgaaa ctggctgaat ctgcagatga ccctcagagt
     6061 caatggtata tttttaatga ttttagcata tcgcccgttt cgccacaaga atcggtatgg
     6121 ttcacattgg attggaaagt tccttgcata ctcttctaca gacacgtaga ggatgactct
     6181 gagtctgcta gtactactag ctcgaccgtc acggaaagtg aagaaaccat tccatcagag
     6241 agcagcagcg gctcgccaac aaatctatcc aacccttttc tagaggaaat tgtgagtcct
     6301 atgctaggca acttaagtgc agacgccacg ctgcagcccc tgcagtcaga tgaaatgccg
     6361 cagtcgggtg atctggtcgc aatggatgcc gagttcgtta ccctaaatcc agaggaaaat
     6421 gagattcgcc cggatggcaa aacggcgacc attaagccat gccacatgag tgtggcccgg
     6481 atttcatgta tacgaggaca aggcccggct gaaggagttc cattcatgga cgattacata
     6541 tccacccagg agaaggtcgt ggactaccta actcagttct cgggaatcaa gcctggcgac
     6601 ttggacgcta acttcagcaa aaagcgactg actgccttgg agtactcaga acagaaatta
     6661 gaatacctcg ttgacgtagg agtaatattt gtcggacatg gtctgaagaa cgactttagg
     6721 gtcataaaca tctatgtacc gtcggagcag ataatagata cggtgcacct gtttcacatg
     6781 ccacatcatc gcatggtttc cctaagattt ctagcctggc atttcttggg tactaagatt
     6841 cagtccgaga cgcacgactc catcgaagat gcacggacca ctctgcagct ctacaagcac
     6901 tatttgaagc tgcaggagga gaagaagttt gccaacgctc tcaagaatct ttacgaacgc
     6961 ggaaagcagc tgcagtggaa ggttccagag gactgaccgc ggttcgaagg taagcctatc
     7021 cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga
     7081 gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc
     7141 tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     7201 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     7261 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     7321 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     7381 gattatgatc agtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     7441 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     7501 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     7561 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     7621 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     7681 ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc
     7741 cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc
     7801 caggcaggta gatgacgacc atcagggaca gcttcaaggc cagcaaaagg ccaggaaccg
     7861 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7921 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7981 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     8041 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     8101 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     8161 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     8221 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     8281 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     8341 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     8401 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8461 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8521 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8581 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8641 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8701 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8761 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8821 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8881 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8941 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     9001 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     9061 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     9121 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     9181 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     9241 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     9301 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     9361 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     9421 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9481 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9541 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9601 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     9661 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattaattct
     9721 atattctaaa aacacaaatg atacttctaa aaaaaaat
//
