LOCUS       sequence-290500-        8443 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Insect Expression of DmPAN3-FLLWGYHRtoEEEAEAAA-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8443)
  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 8443)
  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-DmPAN3-FLLWGYHRtoEEEAEAAA-dsRNAres_Y
FEATURES             Location/Qualifiers
     source          1..8443
                     /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"
     regulatory      3365..3374
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5728..5769
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5779..5796
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5819..5836)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5869..6003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5893..5912)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5947..5966
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6625..7213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6705..6724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7384..8175,8176..8244))
                     /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     8007..8026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8245..8349)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8417..8435
                     /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 ttgaattcta tgtgttccac atacggcctg ctttcattcg aattctcgga
     3361 caccgccgcc atggacccga tctttttttc cccgaccaac ggcattccca gtgaatcgaa
     3421 gctggctaca tacatgaacc gccaaaacgt ggctacgccc tccggatacg gtttgaacaa
     3481 tggatttagt cttctcaacc tggactcccc gctcaacaag aaaagccaag tgacccccca
     3541 atccccggaa tttatcccca cccgtctcgg atccaccccg aatttttacg ccccgtacca
     3601 taatgccccg atgcagctat cgaacggact gaacggagtg aacggtctaa cggctgctgc
     3661 tgctgcggcc gccgccgctg ctgctgctgc tgctgctgcc gtcccgagca gcaccagttc
     3721 cgccagcagc gcttccgtca cgatttccaa gaccgctaat ggcggagcta gcatgctggc
     3781 tttgcagaaa acggctagca ttgcttcagt tacgattgcc cagcaacaac cgcaacaccc
     3841 gcaaaagcaa cagcaacacc ccccctccgt cggaggcggc gccgtcagcg ctgctagcgc
     3901 tccgatcgct atctcgggag ccccgcccaa cccggctgcg gccccctttg tgtcctccat
     3961 gagcgcccaa acccccttga aaggacgagg agctatgatg cgtcaagagt cgccgaccgc
     4021 tgctatgatc tccggagccg gtccgggcga aaaatcgcca ccgcacggga tgacgccgca
     4081 cggagcatcg cccataccat cagcgctgcc caccagcgtg caccaggaga acgtaggcgg
     4141 caccatatac ttctatccga ctgccaacgc acagaacagc cagcccgtcg tcaattccat
     4201 ggtggtggat ggaacgcatc cggctctcca tggcgtgagt gctgtggcgc cgatgagtgc
     4261 gggcgttccg gcggccatga tgtacacggg gcatgtgtat ccgggaccgt cctccaacgt
     4321 ggtcaccatg cagccaaaga cgctgctgga gtcggccttc tttatgcccg acgagatgcg
     4381 cgccgaagtc ctcgcccgca acgagatctc taacctgatc atggacgcag cggaggctgc
     4441 gcagcacgct ctaccgctgg aggtggagaa ctaccatgcc ctgtatccac tggaaccgcc
     4501 ggcgcagccg ttgcacgcca agctcacgtt ccccgccacc acttataggg ccacgcacaa
     4561 cacgacgggc tacaagtact gcctgcgaag aatacatggt ttccgcctgc agtcgaccaa
     4621 gtgcatgacg ctggtggaga tgtggaagaa gctgcagcac acgaatgtgg tacaactgcg
     4681 tgaggtgttc acgacgaaag catttggtga taactcttta gtattagtgt acgactatca
     4741 tcccggctca caaacattac tggccaaata cttcactccg gcgccagaga ccaacgggta
     4801 cactgatcca ttccaaggcg aggcccgccc cttcagtcat aagagtaaca tgcagcgaac
     4861 gagcaatgga ccgctgctac cggaggccac catctggtcg atcatcatgc agctgaccgc
     4921 cggcttaaaa gccatccacc atgccggtct tgcctgcaag gttctggatc ccacaaagat
     4981 tatagtgacg ggcaagcgtg tacggtttag ctcatgctgc atttcggata taacgcaatt
     5041 cgatccgaat gcatccaatc ccttggcgct ggttaatatg catcagcagg acgatctgac
     5101 cgctttgggt cgcttagtat tagctttggc ctgccgctgt cttcaatccg tgcaacgcga
     5161 caatgtccag tccagcatcg atatggtcac gcgaaactac tccaccgatc tgcgaaactt
     5221 tattgtttac ctcttcacca ccaataatcg tcgctccgtg accgacctga tgccgatgat
     5281 tggtgcccgc gaatatacgc aagaagatgc agaacagagc aaaattgata tgcaggagga
     5341 cgagttggcc aaggagatgg agaacgggcg gttatatcgc attttggtca aacttaatag
     5401 catcaacgag cgacccgact ttaatctgga ctgcactgcg tctgagactg aggatagata
     5461 catgttgaaa ttattccgtg atgctttgtt ccattccgtc actgaggatg gccggccctg
     5521 gctagatcac gcagccattg tacaatgcct caacaagctg gatgctggct ccatagaggc
     5581 ggtgcaactg atgtcccgcg acgagcaatc ggtactcatc gtctcctatg ctgaactcaa
     5641 gaattgtctg gagaacgcct tctccgaact gatgtcaagt gcggccaact gagcggccgc
     5701 tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc
     5761 gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     5821 tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga tccagacatg
     5881 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     5941 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     6001 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     6061 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca gtcgaccgat
     6121 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     6181 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     6241 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     6301 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     6361 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
     6421 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     6481 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     6541 tcagggacag cttcaaggat cgctcgcggc tcttaccagc cagcaaaagg ccaggaaccg
     6601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7681 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7981 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     8041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8101 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8341 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     8401 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgt
//
