LOCUS       sequence-357193-        8962 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 8962)
  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 8962)
  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..8962
                     /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             6213..6254
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6264..6281
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6304..6321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6354..6488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6378..6397)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6432..6451
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7089..7677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7169..7188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7848..8639,8640..8708))
                     /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     8471..8490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8709..8813)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8881..8899
                     /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 atggatgact cgtttttcgg ctttgacacg
     3301 aatctaccgg acgaagatgg tggcgatgga cgcatcgtcg aggcggaata cgatgccctc
     3361 aatgacgaaa cctttggatc cgctataaat ggcgactggg aggaagccca tgagacgatg
     3421 gtacgcctgg gcgggaatgg cgaaagggtg cggaagcgac ccgcggaatc tagtggaaat
     3481 acggattttg ccgatcaggg aaatggtgcc ttcctgcgac acgctaatcc gctatcatcg
     3541 gcgccctctt caagcggctc cactccggcc cctttgaatg cccccttccg acagtctcgt
     3601 cacatagggg actcggactt ggagctaaac atttcctcca tgaagttgga cgacatggac
     3661 ctgtcctctt tcaccactga tagcgaagcc ggtgggctga gcaaccgcat taagctggac
     3721 tccggggtct ggggatcaca gccgtttccc aacaaccaac atttgttccg cgagcctatg
     3781 cgcagcgcct tcaagccaca gcagcagcaa catcagcagc accaaccaca acaaatgaag
     3841 ccaccgcagg aagcgaatgc caactttgca ctgaatccac agccttttcc aaaaattaca
     3901 actctcgagg atatagagcg caatatgatc attcagcagg ccattccgaa acaacagcag
     3961 cagagccagc aacaggcgga acgcaaaatg tttgatgatt ttagtctggg taacaggcag
     4021 caggtgccta caccaaccat gttgcagcaa cagatgttgc agcagcagca tcatcagaaa
     4081 caacagtcgc agcagcagcc gccacaacaa gcgcagcaca aggtaccccc tggcttcttg
     4141 ggcacgccgc acacctctcc accgaggcca aatttgggtt tccccggacc cgagctactg
     4201 cccacgccat cgtcgcagca acaactaaac ggtattggtg gcaaccgagt gccgcccgga
     4261 tttatttacc cgcaaggtct gcccgggaat attccgcaat tgccccagca tcctctaatg
     4321 cagcagcaca tgccgcacaa ctttccactg cccggcggca accagcgacc tctacccaat
     4381 ccacaagctc tgcccacggc cctgaacaat ttcgcgatgc atccaagctt caatgcgatg
     4441 cgcgccgctg gaatccatcc aggggctttt atgcaacctc cccatccgca ccagatgctg
     4501 ccaccgcgaa tgccgccgca cccgctgcaa gctgccacaa gtctattagg tcagcagcca
     4561 ccgaattcca tgtataacat gttcaacatg cgactagtgc aagagatcca gcaaaaccat
     4621 ccgctgcttc aacaggctgc tgttgtacgc caaatgcaac agagcagagc aggatccgtg
     4681 gccagcgatc gtcagagatc gcaggtgcaa cagcagcaac aaggacgacg tccagatggt
     4741 acaggaaatt acccgctgga agagtacgat gaatatgcca acctaatgag cacgcgcgac
     4801 aagcactggc taatcggaat ccagctttcg cagctgaata cggacacgcc gtatatcgac
     4861 gactattact acacggtgtt cagggagcgc aaggcccaac agaacgggca aatgcgcaac
     4921 agtcaggcgc acaaggacaa ccagttgaat catccgctta ctcagccacg cgaccatgcg
     4981 cagcttattc tcgtacagtt gggcaacaag aatggtactc gcaatgggca cggacgtgag
     5041 cgacgcaact cggagaacac caacagcacg tcaggcaaca cgaacaatgg atcaggtggt
     5101 aacaacaaca acctgacagg ctatatattc tcaccactga agttcgagaa ctcactggga
     5161 aagctacagt acggaagtgt gacagctccg cgaaaaatta tcgatgctga cattatgagt
     5221 ggtgaatcaa atgccgctgc tgaagccaac gcaacttctt catcaaaaag ctcgactccg
     5281 ctgcccacct ccgcgagcca tgatgtaaat cccagtagca tgcggaagtc gcgacacatt
     5341 ttgcttctga ttgaaacact gtatcgctat ctgctgaagt tggaggattt ggaaagccca
     5401 gaagtgatgg ccaccattga gctgaaaaag aaaaaggaag ccgaacggat cgcagccctt
     5461 gagcagctgg aaatggccaa taaaacgccc gaagaacggg cagcagaggc ggctaaccca
     5521 cagactatga acccacagct caaaaataaa tttaattacg aagttgaaac taaggcagca
     5581 cttgtaaaca agctaaaagc gggacttgct tttgataaag tgattgctat gatgaacgtc
     5641 agaaaaggta aaattctgat ccgccgtatt atgccgttca tagccgacca gtcgatacga
     5701 tgggtggtct ggtccggagt tttttgcagt ctccagactg ttgtcaagaa agataaagat
     5761 gatgtcgagg gagttcttta tgcactgtat ccggaattca agaagcatgt gaataaggca
     5821 acatttgagt tactcgtctc gatatcttca gcaatcactc ttaacgataa aaagctggca
     5881 ggcatcttct gttcccgatt cggcattctg agccttgtgg ctcttattct gagggctgaa
     5941 gagatttatg ttagccgcac cgatgccacg ctgtcggaag aaaatcgcca gaaatggagg
     6001 gagttccttg cccaagtggc cagctgttta aaccggacca tacagaatca aactattagc
     6061 gccgctatcg aaagcgacgc catccaaccc ttgatggatc atttcgagcg cttcaaggac
     6121 ttgaagcttg acgccttact agccctaatt acggaggctc gccaccagat cgactagggc
     6181 ggccgctcgg tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg
     6241 gtctcgattc tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     6301 cagcctcgac tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag
     6361 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     6421 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     6481 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     6541 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga
     6601 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     6661 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     6721 gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg
     6781 tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc
     6841 accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc
     6901 tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc
     6961 gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac
     7021 gaccatcagg gacagcttca aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7501 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7741 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7801 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7861 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7921 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7981 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8041 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8101 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8161 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8221 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8281 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8341 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8401 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8461 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8521 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8581 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8641 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8701 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8761 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8821 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     8881 aataggcgta tcacgaggcc ctttcgtctt caagaattaa ttctatattc taaaaacaca
     8941 aatgatactt ctaaaaaaaa at
//
