LOCUS       sequence-362939-        8835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmPumilio.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8835)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8835)
  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"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmPumilio_B
FEATURES             Location/Qualifiers
     source          1..8835
                     /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             6086..6127
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6137..6154
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6177..6194)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6227..6361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6251..6270)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6305..6324
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6962..7550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7042..7061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7721..8512,8513..8581))
                     /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     8344..8363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8582..8686)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8754..8772
                     /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 gcttgatggt ggttttagaa actgccagcg cccttttggg cggcccgtat
     3301 gcccagggcg ctcccgccct gaaaatggtg cagaaacgct acattggatt acatcactgg
     3361 ctgggaccaa tccgctccaa ggagctaaag gagcatattg taagtgacga cgtgctttcg
     3421 ctcgctttta accacaatca acaactgttt cgctcgcaga atccgggcct agcagcagtt
     3481 gccacaaatg cagcggccgc agcagcagcc gcagcagctg ccacatcggc agcgagtgct
     3541 gcggcagcgg tgggcgcacc acccgttccc aacggatcgc tgcagcagtc gcagcagcaa
     3601 cagcagcagc agcaacaaca gcagcagcaa caacagatgc acatggcggc cgcgtcgcaa
     3661 caatttttgg ccgcccagca gcaggcgcaa aatgcggcct atgccgccca acaggccacg
     3721 tcctacgtca tcaatccggg ccaggaggct gccccgtata tgggcatgat tgccgccgcc
     3781 cagatgccgt actatggcgt agcaccatgg ggcatgtatc cgggcaatct gattccgcaa
     3841 cagggaacgc agccgcgccg ccccctcacc ccctcgcagc agggtgccga gaatcagccg
     3901 tatcaggtca tcccggcatt cctcgatcac acgggctcct tgctgatggg aggacctcgc
     3961 accgggacgc cgatgcgtct ggttagcccc gcccccgttc tggtgccccc gggcgctacc
     4021 cgtgccggcc ccccgccccc gcagggccca cagctgtatc agccgcagcc gcagacggcc
     4081 caacagaatc tctactcgca gcagaatgga tccagtgtcg gaggcctcgc cttgaacacg
     4141 agctcgttga cgggtcgccg cgactccttc gaccgcagca cctccgcctt cagtccctcg
     4201 accatggact acaccagcaa cggtgtggca gcggccgcca atgcggtgaa cagcacagtg
     4261 gcccaggcag cagcagctgc cgcagcagcc gccgcagcgc gtggcaagtg gccgggagcg
     4321 atgtcgggag cggccagtgg agcctacgga gccctgggag cgggcaatgc ttcggccagt
     4381 cccctgggcg caccaatcac gccgccgcca tcggcgcaat cctgtctcct gggcagtcgg
     4441 gcacctggag ccgagtcccg ccagcggcag cagcaacaac agcagctggc cgccgttggt
     4501 ctgccggcga ctgcagcagc tgctcaggca gcggtggccg cggctgccaa caatatgttc
     4561 ggatccaaca gctcgatctt ctcgaatccc ctggccattc cgggtaccgc agctgtggca
     4621 gctgcagcgg cagcagcagc ggccgccaac tcgcgtcagg tggctgccgc ggcagcggca
     4681 gcagcggctg tggcagcagc agccggcgga gtgggaggtg ccccacagcc aggaagatct
     4741 cgccttctcg aagatttccg caaccagcgg tatccaaatc ttcagctacg cgatctcgct
     4801 aaccacattg tggagttctc acaggatcag cacggctcgc ggtttatcca acagaagttg
     4861 gagcgggcca ccgccgccga gaagcaaatg gtgttcagcg agatcctggc ggcagcctat
     4921 agcctgatga ccgatgtctt tggcaactat gtcatccaga agttctttga gttcggcact
     4981 cccgagcaga agaacacgct gggcatgcag gtcaagggtc atgtgctgca gctggcgctg
     5041 caaatgtatg gctgccgagt gattcagaag gctctggaga gcatctcgcc ggagcagcag
     5101 caggaaatcg tccacgaact ggacggacat gtgctgaaat gcgtcaagga tcagaatggc
     5161 aatcatgtgg tgcaaaagtg cattgagtgc gtggaccccg tggcgctgca gttcatcatc
     5221 aatgctttca agggtcaggt ttactcgcta agcacccatc cgtatggatg ccgggtgatc
     5281 cagagaatcc ttgagcattg cactgccgaa cagaccacgc ccattttgga cgaactgcat
     5341 gagcacaccg aacagttgat tcaggaccaa tatggcaact atgttattca gcatgtgctt
     5401 gaacacggca agcaggagga taagtcgatt cttatcaaca gcgtgcgcgg caaagttctg
     5461 gtgctatcac agcacaagtt cgcctcaaac gttgtggaga aatgtgttac ccatgccact
     5521 cgcggagaac gcactggtct catagacgag gtctgcacct tcaacgacaa cgcgttgcac
     5581 gtgatgatga aggatcagta tgccaactat gtggtccaaa aaatgatcga tgtatcggag
     5641 ccgacgcagc tcaagaagct gatgaccaag atccggcccc acatggccgc cttgcgcaag
     5701 tacacctacg gcaagcacat caatgccaag ttggagaagt actacatgaa gataaccaat
     5761 cccattacgg tgggcacagg agctggagga gtgccggcag cctcgtcggc ggccgcagtc
     5821 agcagtggtg ccacctcggc atcggtaacc gcctgcacca gtggcagcag caccaccacg
     5881 accagcacta ccaacagcct ggcctcaccc accatttgtt cggtgcagga gaacggcagc
     5941 gccatggttg tggagccctc ctccccggac gcctccgagt cctcgtcctc ggtggtgtca
     6001 ggcgctgtca acagcagctt gggtcccatt ggacccccga ccaacggcaa cgttgtgctg
     6061 taatctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     6121 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     6181 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     6241 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     6301 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     6361 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     6421 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgaccgatgc
     6481 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     6541 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     6601 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     6661 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     6721 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     6781 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     6841 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     6901 agggacagct tcaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6961 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7021 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7081 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7141 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7201 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7261 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7321 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7381 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7441 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7501 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7561 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7621 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7681 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7741 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7801 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7861 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7921 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7981 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     8041 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8101 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8161 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8221 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8281 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     8341 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8401 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8461 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8521 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8581 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8641 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8701 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8761 gtatcacgag gccctttcgt cttcaagaat taattctata ttctaaaaac acaaatgata
     8821 cttctaaaaa aaaat
//
