LOCUS       sequence-289138-        8987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmHPat-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8987)
  AUTHORS   Haas G, Braun JE, Igreja C, Tritschler F, Nishihara T, Izaurralde E
  TITLE     HPat provides a link between deadenylation and decapping in metazoa.
  JOURNAL   J Cell Biol. 2010 Apr 19. 189(2):289-302.
  PUBMED    20404111
REFERENCE   2  (bases 1 to 8987)
  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"; journalName: "J Cell 
            Biol. 2010 Apr 19. 189(2):289-302."
COMMENT     Sequence Label: pAc5.1B-EGFP-DmHPat-dsRNAres_J
FEATURES             Location/Qualifiers
     source          1..8987
                     /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             6272..6313
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6323..6340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6363..6380)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6413..6547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6437..6456)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6491..6510
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7169..7757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7249..7268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7928..8719,8720..8788))
                     /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     8551..8570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8789..8893)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8961..8979
                     /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 ggatgactcg tttttcggct ttgacacgaa
     3361 tctaccggac gaagatggtg gcgatggacg catcgtcgag gcggaatacg atgccctcaa
     3421 tgacgaaacc tttggatccg ctataaatgg cgactgggag gaagcccatg agacgatggt
     3481 acgcctgggc gggaatggcg aaagggtgcg gaagcgaccc gcggaatcta gtggaaatac
     3541 ggattttgcc gatcagggaa atggtgcctt cctgcgacac gctaatccgc tatcatcggc
     3601 gccctcttca agcggctcca ctccggcccc tttgaatgcc cccttccgac agtctcgtca
     3661 cataggggac tcggacttgg agctaaacat ttcctccatg aagttggacg acatggacct
     3721 gtcctctttc accactgata gcgaagccgg tgggctgagc aaccgcatta agctggactc
     3781 cggggtctgg ggatcacagc cgtttcccaa caaccaacat ttgttccgcg agcctatgcg
     3841 cagcgccttc aagccacagc agcagcaaca tcagcagcac caaccacaac aaatgaagcc
     3901 accgcaggaa gcgaatgcca actttgcact gaatccacag ccttttccaa aaattacaac
     3961 tctcgaggat atagagcgca atatgatcat tcagcaggcc attccgaaac aacagcagca
     4021 gagccagcaa caggcggaac gcaaaatgtt tgatgatttt agtctgggta acaggcagca
     4081 ggtgcctaca ccaaccatgt tgcagcaaca gatgttgcag cagcagcatc atcagaaaca
     4141 acagtcgcag cagcagccgc cacaacaagc gcagcacaag gtaccccctg gcttcttggg
     4201 cacgccgcac acctctccac cgaggccaaa tttgggtttc cccggacccg agctactgcc
     4261 cacgccatcg tcgcagcaac aactaaacgg tattggtggc aaccgagtgc cgcccggatt
     4321 tatttacccg caaggtctgc ccgggaatat tccgcaattg ccccagcatc ctctaatgca
     4381 gcagcacatg ccgcacaact ttccactgcc cggcggcaac cagcgacctc tacccaatcc
     4441 acaagctctg cccacggccc tgaacaattt cgcgatgcat ccaagcttca atgcgatgcg
     4501 cgccgctgga atccatccag gggcttttat gcaacctccc catccgcacc agatgctgcc
     4561 accgcgaatg ccgccgcacc cgctgcaagc tgccacaagt ctattaggtc agcagccacc
     4621 gaattccatg tataacatgt tcaacatgcg actagtgcaa gagatccagc aaaaccatcc
     4681 gctgcttcaa caggctgctg ttgtacgcca aatgcaacag agcagagcag gatccgtggc
     4741 cagcgatcgt cagagatcgc aggcgcaaca gcagcaacaa ggacgacgtc cagatggtac
     4801 aggaaattac ccgctggaag agtacgatga atatgccaac ctaatgagca cgcgcgacaa
     4861 gcactggcta atcggaatcc agctttcgca gctgaatacg gacacgccgt atatcgacga
     4921 ctattactac acggtgttca gggagcgcaa ggcccaacag aacgggcaaa tgcgcaacag
     4981 tcaggcgcac aaggacaacc agttgaatca tccgcttact cagccacgcg gccatgcgca
     5041 gcttattctc gtacagttgg gcaacaagaa tggtactcgc aatgggcacg gacgtgagcg
     5101 acgcaactcg gagaacacca acagcacgtc aggcaacacg aacaatggat caggtggtaa
     5161 caacaacaac ctgacaggct atatattctc accactgaag ttcgagaact cactgggaaa
     5221 gctacagtac ggaagtgtga cagctccgcg aaaaattatc gatgctgaca ttatgagtgg
     5281 tgaatcaaat gccgctgctg aagccaacgc aacttcttca tcaaaaagct cgactccgct
     5341 gcccacctcc gcgagccatg atgtaaatcc cagtagcatg cggaagtcgc gacacatttt
     5401 gcttctgatt gaaacactgt atcgctatct gctgaagttg gaggatttgg aaagcccaga
     5461 agtgatggcc accattgagc tgaaaaagaa aaaggaagcc gaacggatcg cagcccttga
     5521 gcagctggaa atggccaata aaacgcccga agaacgggca gcagaggcgg ctaacccaca
     5581 gactatgaac ccacagctca aaaataaatt taattacgaa gttgaaacta aggcagcact
     5641 tgtaaacaag ctaaaagcgg gacttgcttt tgataaagtg attgctatga tgaacgtcag
     5701 aaaaggtaaa attctgatcc gccgtattat gccgttcata gccgaccagt cgatacgatg
     5761 ggtggtctgg tccggagttt tttgcagtct ccagactgtt gtcaagaaag ataaagatga
     5821 tgtcgaggga gttctttatg cactgtatcc ggaattcaag aagcatgtga ataaggcaac
     5881 atttgagtta ctcgtctcga tatcttcagc aatcactctt aacgataaaa agctggcagg
     5941 catcttctgt tcccgattcg gcattctgag ccttgtggct cttattctga gggctgaaga
     6001 gatttatgtt agccgcaccg atgccacgct gtcggaagaa aatcgccaga aatggaggga
     6061 gttccttgcc caagtggcca gctgtttaaa ccggaccata cagaatcaaa ctattagcgc
     6121 cgctatcgaa agcgacgcca tccaaccctt gatggatcat ttcgagcgct tcaaggactt
     6181 gaagcttgac gccttactag ccctaattac ggaggctcgc caccagatcg actagggcgg
     6241 ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg
     6301 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     6361 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     6421 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6481 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6541 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     6601 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     6661 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     6721 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     6781 cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt
     6841 cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca
     6901 ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct
     6961 acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg
     7021 cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg
     7081 accatcaggg acagcttcaa ggatcgctcg cggctcttac cagccagcaa aaggccagga
     7141 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7201 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7261 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7321 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     7381 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7441 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7501 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7561 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7621 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7681 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7741 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7801 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7861 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7921 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7981 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8041 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8101 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8161 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8221 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8281 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8341 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8401 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8461 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8521 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     8581 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8641 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8701 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8761 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8821 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8881 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     8941 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgt
//
