LOCUS       sequence-358323-        7969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGIGYF_1-640.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7969)
  AUTHORS   Ruscica V, Bawankar P, Peter D, Helms S, Igreja C, Izaurralde E
  TITLE     Direct role for the Drosophila GIGYF protein in 4EHP-mediated mRNA 
            repression.
  JOURNAL   Nucleic Acids Res. 2019 Jul 26;47(13):7035-7048. doi: 
            10.1093/nar/gkz429.
  PUBMED    31114929
REFERENCE   2  (bases 1 to 7969)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2019-07-26- 26"; volume: "47"; issue: 
            "13"; pages: "7035-7048"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmGIGYF_1-640_AH
FEATURES             Location/Qualifiers
     source          1..7969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /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(2630..2651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2891..2910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3238..3259
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5275..5316
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5326..5343
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5366..5383)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5416..5550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5440..5459)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5494..5513
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6151..6739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6231..6250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6910..7701,7702..7770))
                     /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     7533..7552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7771..7875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7943..7961
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 catcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2641 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2701 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2761 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2821 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2881 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2941 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3001 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3061 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3121 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3181 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3241 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3301 gccaagcttg aattctatga cagattcaat gaaatttggc ccggaatggt tgcgcaatat
     3361 gtcagccgag ccttcgggct ctcccagtac ctacaacgtt ggtaatgcag ctcaaaacaa
     3421 ttcgattgga gggcacaacc tgggaaacaa cacggcggca tccgcttccc gtaatctatt
     3481 tccagaatac cggtacggac gcgaggaaat gctgtccttg ttcgatcgga actgcctact
     3541 gccgcagatc ctgccatcgt tcaaaaagct cttcgtggaa aaggttcagt gccctcttgc
     3601 actgacaccg agctccgagg aggacatcaa ccaaaactca cttggcaata actctcgtcc
     3661 tgcctggttg cagcgctcgc ccagtggatt tggtagtgcc tcccgaggct ctggacgtgg
     3721 tggaaccgtt gaccggggac ggatgcgcgg aaaatcggcc tatcatccga tataccaacg
     3781 cccaagcggt ctatatgatg aaagcttatc ggttatatca aaggccgaac gcacatggag
     3841 cgaccgcaac ggaactggag attctgcggc aaccaccacc tccactagtg gctctggtgc
     3901 tctagattgg aatggaactc caagctcaag tcctcgaaaa gattattcta ataatcatcg
     3961 caacttggaa aactggcgac gaacacgtaa cgaagatgga tccggtgatg gtccatcctc
     4021 cagcggttcc atgagcggac ccgatattgc tggttggcgg agcggtgtcg tcggtggaag
     4081 tacaaacgca ggttttggta ccaatagcca tcggtgggga agatctacaa gctggcgtga
     4141 tgaagatcct agcgtcgaca atgcagctag tttacaacgc tcaatttcaa cagttgcgac
     4201 tttaagtaca gatcggactg gaaacaataa aggatctgga atcggagctg cagagggagt
     4261 agggagcata tcgcacccac ggttgtcgag ttctaaaata tctcaattat ggactgttaa
     4321 taatgcagcc ggtgttgatg ccgacgaaaa tctacccgaa tgggcaattg aaaatccatc
     4381 gaaattgggt ggcagttttg atgctagtgg agcctttcat ggagatactg atctaaaacc
     4441 tatcaaaagt tcgcataata ctttaaaaac caaaagctta gactcctatg acgatgtaaa
     4501 aagaccgaaa agtaaagacc tgtctgatcc agatagtgga aacgatataa ctccagaaac
     4561 ctcattaacg aaagatacca atacaacagc tgtgcaagaa gaagttgaaa gcagtttatc
     4621 tccaataagc tctacaacga caaaggaagt gatccatgga gatatttcag atcggattaa
     4681 agaggtcgcc gatgaagtag aaaaacttat aatggatgat gatcataaaa tctcttcgaa
     4741 tcaaagccaa catcaaaatc atgacgggtt tacagctgca cttccaagaa tggcagacat
     4801 tgagataaac gtaaaaccga gtgttacagc agtgcatcgg caagcacctt ccaccatgcc
     4861 tatacaaatt acccctacaa ttacagatgt cgctccacca tcccatgcag ttgtgtcgtt
     4921 ttcggatcat gagacaatgc aacaccataa catgcaccac ctgccacaat ttccaatgat
     4981 tcctacaccg catatcatca ctccaaatct aaacgaattg tggttttacc gggatccgca
     5041 ggcaaatgtt caggggccat ttagtgccgt tgagatgacg gaatggtatc gcgctggcta
     5101 ctttaatgag aacctgtttg tacgccggta ctctgataat aggtttagac cgctgggaga
     5161 gcttataaaa ttttgtcatg gtaacatgcc atttacgcac agtcacctac ttccatctcc
     5221 gatagaacta gagaactgag cggccgctcg agtctagagg gcccgcggtt cgaaggtaag
     5281 cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac
     5341 cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg agctcgctga
     5401 tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg acaaaccaca
     5461 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     5521 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     5581 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     5641 atggctgatt atgatcagtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     5701 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     5761 actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg
     5821 gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca
     5881 agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg
     5941 catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag gctggatggc
     6001 cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat
     6061 gctgtccagg caggtagatg acgaccatca gggacagctt caaggccagc aaaaggccag
     6121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6781 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6841 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6901 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7021 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7081 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7141 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7201 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     7261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7321 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7441 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7501 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     7561 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7621 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7741 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7801 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7861 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7921 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgt
//
