LOCUS       sequence-333415-        6619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmMe31B_268-459.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6619)
  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 6619)
  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-DmMe31B_268-459_D
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..80
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        144..2653
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2673..2675,2676..2678,2679..3389)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2673..2675,2676..2678,2679..3389)
                     /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(2718..2739)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2979..2998)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3326..3347
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4013..4054
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4064..4081
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4104..4121)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4154..4288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4178..4197)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4232..4251
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4889..5477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4969..4988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5648..6439,6440..6508))
                     /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     6271..6290
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6509..6613)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
       61 aaataggcgt atcacgaggc cctttcgtct tcaagaatta attctatatt ctaaaaacac
      121 aaatgatact tctaaaaaaa aatcatgaat ggcatcaact ctgaatcaaa tctttgcaga
      181 tgcacctact tctcatttcc actgtcacat catttttcca gatctcgctg cctgttatgt
      241 ggcccacaaa ccaagacacg ttttatggcc attaaagctg gctgatcgtc gccaaacacc
      301 aaatacataa tgaatatgta cacattcgag aaagaagcga tcaaagaagc gtcttcgggc
      361 ggagtaggag aatgcggagg agaaggagaa cgagctgatc tagtatctct ccacaatcca
      421 atgccaactg accaactggc catattcgga gcaatttgaa gccaatttcc atcgcctggc
      481 gatcgctcca ttcttggcta tatgtttttc accgttaccc ggggccattt tcaaagactc
      541 gtcggcaaga taagattgtg tcactcgctg tctctcttca tttgtcgaag aatgctgagg
      601 aatttcgcga tgacgtcggc gagtattttg aagaatgaga ataatttgta tttatacgaa
      661 aatcagttag tggaattttc tacaaaaaca tgttatctat agataatttt gttgcaaaat
      721 atgttgacta tgacaaagat tgtatgtata tacctttaat gtattctcat tttcttatgt
      781 atttataatg gcaatgatga tactgatgat attttaagat gatgccagac caaaaggctt
      841 gaatttctgc gtcttttgcc gaacgcagtg catgtgcaat tgttgttttt tggaatattc
      901 aattttcgga ctgtccgctt tgatttcagt ttcttggctt attcaaaaag caaagtaaag
      961 ccaaaaaagc gagatggcaa taccaaatgc ggcaaaacgg tagtggaagg aaaggggtgc
     1021 ggggcagcgg aaggaagggt ggggcggggc gtggcggggt ctgtggctgg gcgcgacgtc
     1081 accgacgttg gagccactcc tttgaccatg tgtgcgtgtg tgtattattc gtgtctcgcc
     1141 actcgccggt tgtttttttc tttttatgct gcgctctctc tagcgccatc tcgcttacgc
     1201 atgctcaacg caccgcatgt tgccgtttcc ttttatgcgt cattttggct cgaaataggc
     1261 aattatttaa acaaagatta gtcaacgaaa acgctaaaat aaataagtct acaatatggt
     1321 tacttattgc catgtgtgtg cagccaacga tagcaacaaa agcaacaaca caggtggctt
     1381 tccctctttc actttttgtt tgcaagccgc gtgcgagcaa gacggcacga ccggcaaacg
     1441 caattacgct gacaaagagc agacgaagtt ttggcgaaaa acatcaaggc gcctgatacg
     1501 aatgcatttg caataacaat tgcgatattt aatattgttt atgaagctgt ttgacttcaa
     1561 aacacacaaa aaaaaaaata aaacaaatta tttgaaagag aattaggaat cggacgctta
     1621 tcgttagggt aacaacaaga aatgcttact gagtcacagc ctctggaaaa ctgccgcaag
     1681 ccagagagag agagaaaaag agggagagca gcttagaccg catgtgcttg tgtgtgaggc
     1741 gtctctctct tcgtctctgt tgcgcaaacg catagactgc actgaaaaaa tcgattacct
     1801 attttttatg aatgaatatt tgcactatta ctattcaaaa ctattaagat agcaatcaca
     1861 ttcaatagcc aaatactata ccacctgagc gatgcaacga aatgatcaat ttgagcaaaa
     1921 atgctgcata tttaggacgg catcattata gaaatgcttc ttgctgtgta cttttctctc
     1981 gtctggcagc tgtttcgccg ttattgttaa aaccggctta agttaggtgt gttttctacg
     2041 actagtgaat gccctactag aagatgtgtg ttgcacaaaa tgtccctgga ataaccaatt
     2101 tgaagtgcag atagcagtaa acgtaagcta atatgaatat tatttaactg taatgtttta
     2161 atatcgctgg acattactaa taaacccact ataaacacat gtacatatgt atgttttggc
     2221 atacaatgag tagttgggga aaaaatgtgt aaaagcaccg tgaccatcac agcataaaga
     2281 taaccagctg aagtatcgaa tatgagtaac ccccaaattg aatcacatgc cgcaactgat
     2341 aggacccatg gaagtacact cttcatggcg atatacaaga cacacacaag cacgaacacc
     2401 cagttgcgga ggaaattctc cgtaaatgaa aacccaatcg gcgaacaatt catacccata
     2461 tatggtaaaa gttttgaacg cgacttgaga gcggagagca ttgcggctga taaggtttta
     2521 gcgctaagcg ggctttataa aacgggctgc gggaccagtt ttcatatcac taccgtttga
     2581 gttcttgtgc tgtgtggata ctcctcccga cacaaagccg ctccatcagc cagcagtcgt
     2641 ctaatccaga gaccccggat cggggtacca tcatggtgag caagggcgag gagctgttca
     2701 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     2761 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     2821 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     2881 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2941 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     3001 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3061 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3121 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     3181 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3241 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     3301 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3361 tcactctcgg catggacgag ctgtacaagc caagcttgaa gggtgtcact cagtactatg
     3421 cattcgtaca ggagcgccag aaagtacact gcttgaatac gctcttttcg aagctgcaga
     3481 tcaatcagtc tatcatattc tgcaattcca cgcaacgtgt cgagctattg gccaaaaaga
     3541 tcacagagct tggatattgc tgttactata tacacgctaa gatggcccag gcacacagaa
     3601 atcgagtatt ccacgatttc cgccaaggac tctgtcgcaa tctggtgtgc tccgacctgt
     3661 tcacccgtgg tatcgacgtg caggccgtga atgtagtaat caatttcgat ttcccacgaa
     3721 tggcagagac atacttgcat cgaattggtc gctcaggacg tttcggtcat ttgggtattg
     3781 ctattaatct gataacctac gaggatcggt ttgatctgca tcggattgaa aaagaacttg
     3841 gaaccgaaat aaagcctatt ccgaaggtca tagaccccgc cctttatgtc gcaaacgttg
     3901 gcgcatccgt tggcgatact tgcaacaaca gtgatctgaa taattctgcg aacgaggagg
     3961 gcaacgttag caaataagcg gccgctcgag tctagagggc ccgcggttcg aaggtaagcc
     4021 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     4081 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     4141 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     4201 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4261 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4321 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     4381 ggctgattat gatcagtcga ccgatgccct tgagagcctt caacccagtc agctccttcc
     4441 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
     4501 tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga
     4561 gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag
     4621 ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca
     4681 tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct
     4741 tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc
     4801 tgtccaggca ggtagatgac gaccatcagg gacagcttca aggccagcaa aaggccagga
     4861 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4921 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4981 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5041 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5101 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5161 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5221 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5281 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5341 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5401 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5461 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5521 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5581 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5641 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5701 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5761 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5821 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5881 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5941 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6001 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6061 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6121 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6181 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6241 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     6301 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6361 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6421 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6481 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6541 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6601 taggggttcc gcgcacatt
//
