LOCUS       sequence-148403-        7974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-tagged DmNot10 in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7974)
  AUTHORS   Bawankar P, Loh B, Wohlbold L, Schmidt S, Izaurralde E
  TITLE     NOT10 and C2orf29/NOT11 form a conserved module of the CCR4-NOT 
            complex that docks onto the NOT1 N-terminal domain.
  JOURNAL   RNA Biol. 2013 Feb;10(2):228-44. doi: 10.4161/rna.23018. Epub 2013 
            Jan 9.
  PUBMED    23303381
REFERENCE   2  (bases 1 to 7974)
  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: "10.4161/rna.23018";
            journalName: "RNA Biol"; date: "2013-02"; volume: "10"; issue: "2"; 
            pages: "228-44"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmNot10
FEATURES             Location/Qualifiers
     source          1..7974
                     /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             5259..5300
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5310..5327
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5350..5367)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5400..5534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5424..5443)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5478..5497
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6156..6744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6236..6255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6915..7706,7707..7775))
                     /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     7538..7557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7776..7880)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7948..7966
                     /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 ggactcggcg gaatccccca ccaaaacgca
     3361 ggccggcgag gacgagaact acagtctgct gtgccaggcg cacgagcagt tcaacaactc
     3421 ggagtttgat cgctgcttgg aactgctcca ggagttggag acgcgcggcg agagcagtgg
     3481 tcctgtcctg cggcacaacc gggcggtggt cagctactac aaaaccgggt gcacccaaca
     3541 ctccgtcctg ctgaaagaac tggaggctct aaccgccgat gcagatgcgc ccggagacgt
     3601 gtccagcggg ctgagcctta agcagggagc tgccgccgcc actgttgctc gttacaaccg
     3661 ggcggtcatc tattatcacc gccatatgtt cggcacggca ctggagaaac tagcaccact
     3721 agtggctcgt ctggaagccc tggagaaggc aatggcagcg ctggtggcca cactacagct
     3781 acagctgctg ctggccacca atcaactaaa ccgagccgag gcctttctgg attacttgca
     3841 gtacaagctt aacctggtgg ccacagcgcc tagcagcaat gcggccgaag aggcagctgt
     3901 tgtcggcaca gcacctccgt cagcagcgac gaacagttca gtggtggcct caccaggagg
     3961 agcagctgtg gagggagctg tgcccggtct gggcggtagt ctgcagctct tgcagctaat
     4021 aacgctcgtg cttaaccgaa aaccggtggt tattcaggaa gacggcacgc cggagtacgc
     4081 tgctctgaag gctcagcagt actacatcat gaaggatttt cagatggccg ccaagcagct
     4141 tatgcgaatc aataacgagt gtacacaagc gggcactata accccccagc tgagcacgtg
     4201 cattgccaac aatatgggag tgatacactt aagagtgcgt cactatgcca tcgccgccaa
     4261 gttcttccaa aatgcgctca actttgacca gcagctggcg cgaaatttgc gccagagtac
     4321 gctgcaaacc atgagttcgg cacggtcctg cgagatcctt tacaacctcg gtgtggccat
     4381 gttgcaccta cgacgcccca aggaggcgtt tcagtgcttt ctggttcccg ttaagcaatt
     4441 ccacagcaat ccacgtcttt ggctgcgcat ggctgaggcc tgcatcatgg agcatgaggc
     4501 gaaactagtg gaggaggagc gccagtcgca gtcagaaacg ccatcaacaa agccatacgc
     4561 tcctcaatcg gcgggagttc cagagcccac gctggaattc gctgtcttgt gtctacgtag
     4621 cgcgttaacg cttacgcagc actacaagac gaggtttcac atggcggcgg tctcctcgga
     4681 ggatgtggaa gcacccgaac ccaaggaccc aactcaggag agttggcgtc atccccagga
     4741 caataacttc tgcaatccct cgaagccagt cagcctagag tctttggaaa acatgatggc
     4801 tgccatatat gctgctcaca gcttcgtctc gctgcgccta ggcgatcatg tgaccgcgct
     4861 cgagatgtcg gaaaaactgc tggcctgtga acgcttgtcc gatgcacaca aattgctagg
     4921 tcacatgtac gctggcgagg cgttgatgct tatggacaag gccagcgagg cacgcgacca
     4981 tttggaccct acttttgtgg gcacactaaa cgccttcgat tttgagacgc gtgactggca
     5041 actgaagtcg gtggacgccg ctcagaacgt tgtgcggtat aatctggccg ttgccatggc
     5101 tctacagaac gacctgcctc aagctaaagc actcctggcc aatttaactc attcgctggt
     5161 tgccaacaag gcgcttgctt tgcgacgctt catggacctc aaaatgggtc ccgttcctgg
     5221 cgggacgccg agttagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc
     5281 tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc
     5341 gctgatcagc ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg
     5401 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     5461 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     5521 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     5581 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     5641 agtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     5701 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     5761 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
     5821 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
     5881 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg
     5941 ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt
     6001 cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta
     6061 gatgacgacc atcagggaca gcttcaagga tcgctcgcgg ctcttaccag ccagcaaaag
     6121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     6361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6901 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7021 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7201 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     7261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7501 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     7561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7921 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgt
//
