LOCUS       sequence-290486-        9623 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Insect Expression of DmPAN2-del975-1031-GSSG-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9623)
  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 9623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-del975-1031GSSG-dsRNAres_Y
FEATURES             Location/Qualifiers
     source          1..9623
                     /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             6908..6949
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6959..6976
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6999..7016)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7049..7183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7073..7092)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7127..7146
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7805..8393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7885..7904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8564..9355,9356..9424))
                     /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     9187..9206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9425..9529)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9597..9615
                     /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 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 tagggttcct
     4981 cttcaacatg ctggataaaa 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 ggcagcagcg gctcgggtga tctggtcgca atggatgccg agttcgttac
     6301 cctaaatcca gaggaaaatg agattcgccc ggatggcaaa acggcgacca ttaagccatg
     6361 ccacatgagt gtggcccgga tttcatgtat acgaggacaa ggcccggctg aaggagttcc
     6421 attcatggac gattacatat ccacccagga gaaggtcgtg gactacctaa ctcagttctc
     6481 gggaatcaag cctggcgact tggacgctaa cttcagcaaa aagcgactga ctgccttgaa
     6541 gtactcatac cagaaattaa aatacctcgt tgacgtagga gtaatatttg tcggacatgg
     6601 tctgaagaac gactttaggg tcataaacat ctatgtaccg tcggagcaga taatagatac
     6661 ggtgcacctg tttcacatgc cacatcatcg catggtttcc ctaagatttc tagcctggca
     6721 tttcttgggt actaagattc agtccgagac gcacgactcc atcgaagatg cacggaccac
     6781 tctgcagctc tacaagcact atttgaagct gcaggaggag aagaagtttg ccaacgctct
     6841 caagaatctt tacgaacgcg gaaagcagct gcagtggaag gttccagagg actgaccgcg
     6901 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     6961 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     7021 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     7081 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     7141 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     7201 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     7261 ctacaaatgt ggtatggctg attatgatca gtcgaccgat gcccttgaga gccttcaacc
     7321 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     7381 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
     7441 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
     7501 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
     7561 ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc
     7621 gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg
     7681 cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat
     7741 cgctcgcggc tcttaccagc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7801 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7921 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7981 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     8041 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8101 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8161 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8221 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     8281 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8341 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8401 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8461 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8521 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8581 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8641 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8701 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8761 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8821 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     8881 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8941 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     9001 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     9061 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9121 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     9181 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9241 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9301 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9361 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9421 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9481 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9541 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     9601 ggcgtatcac gaggcccttt cgt
//
