LOCUS       sequence-441309-        6987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGIGYF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6987)
  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 6987)
  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-lambdaN-HA-DmGIGYF_AH
FEATURES             Location/Qualifiers
     source          1..6987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1501..1692
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1686..1708)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1794..1933
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1849..1868
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1948..1965)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2119..2707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2199..2218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2878..3669,3670..3738))
                     /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     3501..3520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3739..3843)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3911..3929
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(3956..3961,3962..3978,3979..3984)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(4159..4206)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4202..4220
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4482..4505)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(4521..5621)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="METEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     RBS             5628..5650
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5665..5689
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(5689..5708)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5690..5708)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5792..5811)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6017..6094
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(6114..6133)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
       61 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
      121 tccccgcgcg ttggccgatt cattaatgca gctggcacga cagtttcccg actggaaagc
      181 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
      241 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
      301 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
      361 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
      421 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
      481 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg
      541 ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct
      601 cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga
      661 cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat
      721 tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc
      781 atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg
      841 gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact
      901 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg
      961 cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc
     1021 gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc
     1081 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc
     1141 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
     1201 gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc
     1261 atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg
     1321 cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc
     1381 tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc
     1441 tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca
     1501 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta
     1561 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg
     1621 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
     1681 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     1741 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     1801 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     1861 agattgtact gagagtgcac cattgcggtg tgaaataccg cacagatgcg taaggagaaa
     1921 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1981 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2041 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2101 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2161 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2221 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2281 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2341 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2401 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2461 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2521 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     2581 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2641 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2701 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2761 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2821 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2881 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2941 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3001 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3061 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3121 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3181 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3241 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3301 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3361 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3421 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3481 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3541 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3601 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3661 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3721 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3781 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3841 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3901 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct catgtttgac
     3961 agcttatcat cgataagctt taatgcggta gtttatcaca gttaaattgc taacgcagtc
     4021 aggcaccgtg tatgaaatct aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg
     4081 atgctgtagg cataggcttg gttatgccgg tactgccggg cctcttgcgg gatatccgga
     4141 tatagttcct cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta
     4201 tgctagttat tgctcagcgg gtggcagcag ccaactcatt ccgttacggt gtaggttttg
     4261 gtagcgtcgt cgtaggtcca ttcaccgtca acaccgttgt cgttagcgta ctgtttgaaa
     4321 actttttccg cggtagcagc gtcaacagct tcggtggtgg tttcaccttt cagggtttta
     4381 ccgttcagga taagtttgta ctgcatgccg ctagctctag aggatcctgc caggctcggt
     4441 ttcagatgat acagttcgtt catggtatat ttaatcatat ggggcccctg gaacagaact
     4501 tccaggccgc tgccggtacc agtctgcgcg tctttaaggg cttcatcgac agtctgacga
     4561 ccgctggcgg cgttgatcac cgcagtacgc acggcatacc agaaagcgga catctgcggg
     4621 atgttcggca tgatttcacc tttctgggcg ttttccatcg tggcggcaat acgtggatct
     4681 ttcgccaact cttcctcgta agacttcagc gctacggcac ccagcggttt gtctttatta
     4741 accgcttcca gaccttcatc agtcagcaga tagttttcga ggaactcttt cgccagctct
     4801 ttgttcggac tggcggcgtt aatacctgcg ctcagcacgc caacgaacgg tttggatggt
     4861 tgacccttga aggtcggcag taccgttaca ccataattca ctttgctggt gtcgatgttg
     4921 gaccatgccc acgggccgtt gatggtcatc gctgtttcgc ctttattaaa ggcagcttct
     4981 gcgatggagt aatcggtgtc tgcattcatg tgtttgtttt taatcaggtc aaccaggaag
     5041 gtcagacccg ctttcgcgcc agcgttatcc acgcccacgt ctttaatgtc gtacttgccg
     5101 ttttcatact tgaacgcata acccccgtca gcagcaatca gcggccaggt gaagtacggt
     5161 tcttgcaggt tgaacatcag ggcgctctta cctttcgctt tcagttcttt atccagcgcc
     5221 gggatctctt cccaggtttt tggcgggttc ggcagcaggt ctttgttata aatcagcgat
     5281 aacgcttcaa cagcgatcgg gtaagcaatc agcttgccgt tgtaacgtac ggcatcccag
     5341 gtaaacggat acagcttgtc ctggaacgct ttgtccgggg tgatttcagc caacaggcca
     5401 gattgagcgt agccaccaaa gcggtcgtgt gcccagaaga taatgtcagg gccatcgcca
     5461 gttgccgcaa cctgtgggaa tttctcttcc agtttatccg gatgctcaac ggtgacttta
     5521 attccggtat ctttctcgaa tttcttaccg acttcagcga gaccgttata gcctttatcg
     5581 ccgttaatcc agattaccag tttaccttct tcagtttcca tggtatatct ccttcttaaa
     5641 gttaaacaaa attattacta gtggggaatt gttatccgct cacaattccc ctatagtgag
     5701 tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc
     5761 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     5821 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     5881 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
     5941 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     6001 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     6061 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     6121 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     6181 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     6241 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     6301 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     6361 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     6421 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     6481 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     6541 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     6601 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     6661 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     6721 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     6781 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     6841 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     6901 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     6961 caaacaggat tttcgcctgc tggggca
//
