LOCUS       sequence-289136-        8822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmHPat_57-968-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8822)
  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 8822)
  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_57-968-dsRNAres_J
FEATURES             Location/Qualifiers
     source          1..8822
                     /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             6107..6148
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6158..6175
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6198..6215)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6248..6382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6272..6291)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6326..6345
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7004..7592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7084..7103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7763..8554,8555..8623))
                     /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     8386..8405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8624..8728)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8796..8814
                     /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 gggcgaaagg gtgcggaagc gacccgcgga
     3361 atctagtgga aatacggatt ttgccgatca gggaaatggt gccttcctgc gacacgctaa
     3421 tccgctatca tcggcgccct cttcaagcgg ctccactccg gcccctttga atgccccctt
     3481 ccgacagtct cgtcacatag gggactcgga cttggagcta aacatttcct ccatgaagtt
     3541 ggacgacatg gacctgtcct ctttcaccac tgatagcgaa gccggtgggc tgagcaaccg
     3601 cattaagctg gactccgggg tctggggatc acagccgttc cccaacaacc aacatttgtt
     3661 ccgcgagcct atgcgcagcg ccttcaagcc acagcagcag caacatcagc agcaccaacc
     3721 acaacaaatg aagccaccgc aggaagcgaa tgccaacttt gcactgaatc cacagccttt
     3781 tccaaaaatt acaactctcg aggatataga gcgcaatatg atcattcagc aggccattcc
     3841 gaaacaacag cagcagagcc agcaacaggc ggaacgcaaa atgtttgatg attttagtct
     3901 gggtaacagg cagcaggtgc ctacaccaac catgttgcag caacagatgt tgcagcagca
     3961 gcatcatcag aaacaacagt cgcagcagca gccgccacaa caagcgcagc acaaggtacc
     4021 ccctggcttc ttgggcacgc cgcacacctc tccaccgagg ccaaatttgg gtttccccgg
     4081 acccgagcta ctgcccacgc catcgtcgca gcaacaacta aacggtattg gtggcaaccg
     4141 agtgccgccc ggatttattt acccgcaagg tctgcccggg aatattccgc aattgcccca
     4201 gcatcctcta atgcagcagc acatgccgca caactttcca ctgcccggcg gcaaccagcg
     4261 acctctaccc aatccacaag ctctgcccac ggccctgaac aatttcgcga tgcatccaag
     4321 cttcaatgcg atgcgcgccg ctggaatcca tccaggggct tttatgcaac ctccccatcc
     4381 gcaccagatg ctgccaccgc gaatgccgcc gcacccgctg caagctgcca caagtctatt
     4441 aggtcagcag ccaccgaatt ccatgtataa catgttcaac atgcgactag tgcaagagat
     4501 ccagcaaaac catccgctgc ttcaacaggc tgctgttgta cgccaaatgc aacagagcag
     4561 agcaggatcc gtggccagcg atcgtcagag atcgcaggcg caacagcagc aacaaggacg
     4621 acgtccagat ggtacaggaa attacccgct ggaagagtac gatgaatatg ccaacctaat
     4681 gagcacgcgc gacaagcact ggctaatcgg aatccagctt tcgcagctga atacggacac
     4741 gccgtatatc gacgactatt actacacggt gttcagggag cgcaaggccc aacagaacgg
     4801 gcaaatgcgc aacagtcagg cgcacaagga caaccagttg aatcatccgc ttactcagcc
     4861 acgcgaccat gcgcagctta ttctcgtaca gttgggcaac aagaatggta ctcgcaatgg
     4921 gcacggacgt gagcgacgca actcggagaa caccaacagc acgtcaggca acacgaacaa
     4981 tggatcaggt ggtaacaaca acaacctgac aggctatata ttctcaccac tgaagttcga
     5041 gaactcactg ggaaagctac agtacggaag tgtgacagct ccgcgaaaaa ttatcgatgc
     5101 tgacattatg agtggtgaat caaatgccgc tgctgaagcc aacgcaactt cttcatcaaa
     5161 aagctcgact ccgctgccca cctccgcgag ccatgatgta aatcccagta gcatgcggaa
     5221 gtcgcgacac attttgcttc cgattgaaac actgtatcgc tatctgctga agttggagga
     5281 tttggaaagc ccagaagtga tggccaccat tgagctgaaa aagaaaaagg aagccgaacg
     5341 gatcgcagcc cttgagcagc tggaaatggc caataaaacg cccgaagaac gggcagcaga
     5401 ggcggctaac ccacagacta tgaacccaca gctcaaaaat aaatttaatt acgaagttga
     5461 aactaaggca gcacttgtaa acaagctaaa agcgggactt gcttttgata aagtgattgc
     5521 tatgatgaac gtcagaaaag gtaaaattct gatccgccgt attatgccgt tcatagccga
     5581 ccagtcgata cgatgggtgg tctggtccgg agttttttgc agtctccaga ctgttgtcaa
     5641 gaaagataaa gatgatgtcg agggagttct ttatgcactg tatccggaat tcaagaagca
     5701 tgtgaataag gcaacatttg agttactcgt ctcgatatct tcagcaatca ctcttaacga
     5761 taaaaagctg gcaggcatct tctgttcccg attcggcatt ctgagccttg tggctcttat
     5821 tctgagggct gaagagattt atgttagccg caccgatgcc acgctgtcgg aagaaaatcg
     5881 ccagaaatgg agggagttcc ttgcccaagt ggccagctgt ttaaaccgga ccatacagaa
     5941 tcaaactatt agcgccgcta tcgaaagcga cgccatccaa cccttgatgg atcatttcga
     6001 gcgcttcaag gacttgaagc ttgacgcctt actagcccta attacggagg ctcgccacca
     6061 gatcgactag ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc
     6121 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     6181 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     6241 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     6301 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     6361 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     6421 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     6481 ttatgatcag tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     6541 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     6601 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga
     6661 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg
     6721 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg
     6781 ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca
     6841 ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca
     6901 ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc
     6961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     7201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7441 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7561 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7621 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7681 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7741 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7801 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7861 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7921 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7981 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8041 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     8101 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8161 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8221 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8281 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8341 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     8401 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8461 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8521 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8581 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8641 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8701 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8761 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8821 gt
//
