LOCUS       sequence-291266-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmCaf40-V186E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  AUTHORS   Sgromo A, Raisch T, Backhaus C, Keskeny C, Alva V, Weichenrieder O, 
            Izaurralde E
  TITLE     Drosophila Bag-of-marbles directly interacts with the CAF40 subunit 
            of the CCR4-NOT complex to elicit repression of mRNA targets.
  JOURNAL   RNA. 2018 Mar;24(3):381-395. doi: 10.1261/rna.064584.117. Epub 2017 
            Dec 18.
  PUBMED    29255063
REFERENCE   2  (bases 1 to 6972)
  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"; doi: 
            "10.1261/rna.064584.117"; journalName: "RNA"; date: "2018-03"; 
            volume: "24"; issue: "3"; pages: "381-395"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmCaf40-V186E_AG
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..64
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             74..91
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(114..131)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    164..298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(188..207)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     242..261
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(920..1508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1000..1019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1679..2470,2471..2539))
                     /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     2302..2321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2540..2644)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2712..2730
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2794..5306
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(5326..5328,5329..5331,5332..6042)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5326..5328,5329..5331,5332..6042)
                     /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(5371..5392)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5632..5651)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5979..6000
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
       61 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
      121 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
      181 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      241 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      301 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      361 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga ccgatgccct
      421 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
      481 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
      541 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
      601 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
      661 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
      721 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
      781 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
      841 gacagcttca aggatcgctc gcggctctta ccagccagca aaaggccagg aaccgtaaaa
      901 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      961 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1021 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1081 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     1141 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1201 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1261 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1321 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     1381 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1441 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1501 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1561 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1621 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1681 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1741 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1801 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1861 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1921 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1981 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2041 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2101 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2161 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2221 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2281 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2341 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2401 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2461 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2521 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2581 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     2641 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     2701 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaaggaatt aattctatat
     2761 tctaaaaaca caaatgatac ttctaaaaaa aatcatgaat ggcatcaact ctgaatcaaa
     2821 tctttgcaga tgcacctact tctcatttcc actgtcacat catttttcca gatctcgctg
     2881 cctgttatgt ggcccacaaa ccaagacacg ttttatggcc attaaagctg gctgatcgtc
     2941 gccaaacacc aaatacatat caatatgtac attcgagaaa gaagcgatca aagaagcgtc
     3001 ttcgggcgag taggagaatg cggaggagaa ggagaacgag ctgatctagt atctctccac
     3061 aatccaatgc caactgacca actggccata ttcggagcaa tttgaagcca atttccatcg
     3121 cctggcgatc gctccattct tggctatatg tttttcaccg ttcccggggc cattttcaaa
     3181 gactcgtcgg taagataaga ttgtgtcact cgctgtctct cttcatttgt cgaagaatgc
     3241 tgaggaattt cgcgatgacg tcggcgagta ttttgaagaa tgagaataat ttgtatttat
     3301 acgaaaatca gttagtggaa ttttctacaa aaacatgtta tctatagata attttgttgc
     3361 aaaatatgtt gactatgaca aagattgtat gtatatacct ttaatgtatt ctcattttct
     3421 tatgtattta taatggcaat gatgatactg atgatatttt aagatgatgc cagaccacag
     3481 gctgatttct gcgtcttttg ccgaacgcag tgcatgtgcg gttgttgttt tttggaatag
     3541 tttcaatttt cggactgtcc gctttgattt cagtttcttg gcttattcaa aaagcaaagt
     3601 aaagccaaaa aagcgagatg gcaataccaa atgcggcaaa acggtagtgg aaggaaaggg
     3661 gtgcggggca gcggaaggaa gggtggggcg gggcgtggcg gggtctgtgg ctgggcgcga
     3721 cgtcaccgac gttggagcca ctcctttgac catgtgtgcg tgtgtgtatt attcgtgtct
     3781 cgccactcgc cggttgtttt tttcttttta tctcgctctc tctagcgcca tctcgtacgc
     3841 atgctcaacg caccgcatgt tgccgtgtcc tttatgcgtc attttggctc gaaataggca
     3901 attatttaaa caaagattag tcaacgaaaa cgctaaaata aataagtcta caatatggtt
     3961 acttattgcc atgtgtgtgc agccaacgat agcaacaaaa gcaacaacac agtggctttc
     4021 cctctttcac tttttgtttg caagcgcgtg cgagcaagac ggcacgaccg gcaaacgcaa
     4081 ttacgctgac aaagagcaga cgaagttttg gccgaaaaac atcaaggcgc ctgatacgaa
     4141 tgcatttgca ataacaattg cgatatttaa tattgtttat gaagctgttt gacttcaaaa
     4201 cacacaaaaa aaaaaataaa acaaattatt tgaaagagaa ttaggaatcg gacagcttat
     4261 cgttacgggc taacagcaca ccgagacgaa atagcttacc tgacgtcaca gcctctggaa
     4321 gaactgccgc caagcagaga gagagagaaa aagagggaga gcagcttaga ccgcatgtgc
     4381 ttgtgtgtga ggcgtctctc tcttcgtctc ctgtttgcgc aaacgcatag actgcactga
     4441 gaaaatcgat tacctatttt ttatgaatga atatttgcac tattactatt caaaactatt
     4501 aagatagcaa tcacattcaa tagccaaata ctataccacc tgagcgatgc aacgaaatga
     4561 tcaatttgag caaaaatgct gcatatttag gacggcatca ttatagaaat gcttcttgct
     4621 gtgtactttt ctctcgtctg gcagctgttt cgccgttatt gttaaaaccg gcttaagtta
     4681 ggtgtgtttt ctacgactag tgatgcccct actagaagat gtgtgttgca caaatgtccc
     4741 tgaataacca atttgaagtg cagatagcag taaacgtaag ctaatatgaa tattatttaa
     4801 ctgtaatgtt ttaatatcgc tggacattac taataaaccc actataaaca catgtacata
     4861 tgtatgtttt ggcatacaat gagtagttgg ggaaaaaatg tgtaaaagca ccgtgaccat
     4921 cacagcataa agataaccag ctgaagtatc gaatatgagt aacccccaaa ttgaatcaca
     4981 tgccgcaact gataggaccc atggaagtac actcttcatg gcgatataca agacacacac
     5041 aagcacgaac acccagttgc ggaggaaatt ctccgtaaat gaaaacccaa tcggcgaaca
     5101 attcataccc atatatggta aaagttttga acgcgacttg agagcggaga gcattgcggc
     5161 tgataaggtt ttagcgctaa gcgggcttta taaaacgggc tgcgggacca gttttcatat
     5221 cactaccgtt tgagttcttg tgctgtgtgg atactcctcc cgacacaaag ccgctccatc
     5281 agccagcagt cgtctaatcc agagaccccg gatcggggta ccaacatggt gagcaagggc
     5341 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     5401 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     5461 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     5521 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     5581 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     5641 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     5701 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     5761 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     5821 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     5881 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     5941 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     6001 accgccgccg ggatcactct cggcatggac gagctgtaca agccaagctt aattaagatg
     6061 agtgctcaac cgagtccgca tatgaatcct cagcagcagc agcagcagca acagcagcag
     6121 cagcagaccg agcaggagaa ggtgtaccag tggatcaatg agctggccca tccggacacg
     6181 cgtgaaaccg ctctgctcga gctaagcaag aagcgtgaga cggacctggc ccccatgctg
     6241 tggaacagct tcgggaccgc ctgcgccctg ttgcaggaga ttgttaacat ctacccatcg
     6301 ataacgccgc ccactttgac ggcccaccag tcgaaccgcg tgtgcaacgc cctggcgctg
     6361 ctgcagtgcg tcgcctcgca cccggagact cgcacggcct tcctgcaggc ccagataccg
     6421 ctgtacttgt accctttctt gtcgaccacg tccaagacca ggcccttcga gtacttgcgc
     6481 ctgaccagtt tgggcgtgat tggtgctctg gtcaagaccg acgaacagga ggtgatcacc
     6541 tttctgctga ccaccgagat cgtgcccctt tgtctgagca tcatggacag cggatcggag
     6601 ctgagcaaga ctgaagccac tttcatcatc cagaagatcc tgctcgatga gtcgggtctg
     6661 tcgtacatct gccagaccta cgaacgcttt tcgcacgtgg ccatcaccct gggcaaaatg
     6721 gtcatccagc tggcgaagga tccatgcgcc cggctgttga agcatgtggt gcgctgctat
     6781 ctgcgtcttt cggacaatac acgcgctcgc aaggccctgg gacagtgtct gcccgatcag
     6841 ctgcgtgacg gcacctttgc gctgtgcctg caagaggaca agtcgaccaa gcagtggctg
     6901 cagatgctgc tcaagaacct ggaactcggc gccactccgc agcagatcgg catgtcgcca
     6961 ctgggctcct ag
//
