LOCUS       sequence-290025-        9940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmCYFIP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9940)
  AUTHORS   Igreja C, Peter D, Weiler C, Izaurralde E
  TITLE     4E-BPs require non-canonical 4E-binding motifs and a lateral surface
            of eIF4E to repress translation.
  JOURNAL   Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790.
  PUBMED    25179781
REFERENCE   2  (bases 1 to 9940)
  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"; journalName: "Nat 
            Commun."; date: "2014-09-2"; volume: "5:4790. doi"; pages: " 
            10.1038/ncomms5790"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmCYFIP1_T
FEATURES             Location/Qualifiers
     source          1..9940
                     /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             complement(4485..4496)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             7225..7266
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7276..7293
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7316..7333)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7366..7500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7390..7409)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7444..7463
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(8122..8710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8202..8221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8881..9672,9673..9741))
                     /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     9504..9523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9742..9846)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9914..9932
                     /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 ttaatgacgg agaagattac gctagccgac gcgctgtcca acgtggaggt
     3361 gttggacgag ctatccctgc cggacgagca gccctgcatc gaggcgcagc cctgctcgat
     3421 tatctacaag gcaaacttcg atacgaattt cgaggatcgc aatggatttg tcacgggtat
     3481 cgccaagtac atcgaggagg ccaccaccca tgccaacttg aatgtgctcc tggacgaggg
     3541 gcagaaacac gcagtaatgc tctacacctg gcgctgctgc tcgcgcgcca ttccgcagcc
     3601 caagtccaat gagcagccga accgcgtgga gatctacgag aagacggtcg aggtgctagc
     3661 cccggaggtg aacaagctgc tcaacttcat gtacttccaa cgcaaggcca tcgaggcctt
     3721 ctccggcgag gtgaagcgct tgtgccacgt cgagaagcgc aaggacttcg tgtccgaggc
     3781 gtacctattg acgctaggca agtttattaa catgtttgcc gtgctggacg agctgaagaa
     3841 catgaagtcc agcgtgaaga acgattactc cacctacagg cgagctgcgc agttcctcaa
     3901 agtgatgtcg gactcgcaca ctttgcagga gtcgcagaat ctgtccatgt tcctggccac
     3961 gcagaacaag attcgcgaca cggtcaagga cacgctggag aagatcgtgg gctacgagga
     4021 tctgctctca gatgtggtca acatttgcgt gcatatgttc gagacaaaga tgtacttgac
     4081 ccctgaggag aagcacatgc tggtaaaagt tatgggcttt ggcctgttcc ttatggacag
     4141 cgatgcgtgc aatatcaaca aacttgatca aaagaagaag atacggctgg atcgcatcga
     4201 tcgcatcttt aagaacctgg aagtggtgcc cctcttcggc gacatgcaga ttgctccctt
     4261 caactacata aagcgcagca agcacttcga ttccagcaag tggccgctgt ccagctctaa
     4321 cgccatcagc ccacaggcgg atctaatggt gcatttgccg cagatacgcg aggatcacgt
     4381 caagtacatt tcggagctgg cgcgatacac aaacgaggtg accaccaccg tcaaggagaa
     4441 tccctctgat gccgagaacc ggattacggc cgacctggct ctgcgcggcc ttcaattgct
     4501 gtccgagtgg accagtgtgg tcaccgagtt gtactcctgg aagctcctgc atccgactga
     4561 tcatcaccag aacaaggagt gccccgtgga ggctgaggag tacgagcggg ccacccgcta
     4621 caattacacc tcggaagaga agttcgcgct gatcgaggtc atcgccatga taaagggact
     4681 gcaggtgcta atggcgcgca ttgaaaccgt attgtgcgag gctatccggc gtaacatcta
     4741 ttccgagctg caggacttcg tgcagctttc gcttcgtgaa ccgctgcgaa aggccgtaaa
     4801 aaataagaag gatcttatcc ggagcatcat catgtcggtg cgagagacat cggcggactg
     4861 gcaaaagggc tatgagccca ccgacgatcc agtggccaag ggcaagaagg atcccgacgg
     4921 cggcttccgc atccaggtgc ctcgcctcaa tgtgggacct tcctccactc agctatacat
     4981 ggtgcgcacc atgctggagt ctctgattgc cgacaaaagc ggcggcaagc gcactttgcg
     5041 caaggacatc gacggaaact gtttgctgca gatcgacacg ttccacaaga catcgttcta
     5101 ctggagctac ttgctcaact tcagcgacac gcttcagaag tgctgcgatc tctcgcagct
     5161 gtggtatcgc gagttctact tggagatgac catgggtcgt aaggttaaca agtgcttggt
     5221 gcggcatcag cacaacgagg agtgcaagga tctgatcacc atggagaagc gcattcagtt
     5281 tccgattgaa atgtccatgc cctggatcct aaccgatcac attctgcaaa cgaaggagcc
     5341 ctcaatgatg gaatttgtct tgtatcccct ggacttgtac aacgactcgg cgtactacgc
     5401 actcactgtc ttccgaaagc agttcctgta cgacgaagtg gaggccgaag tcaatctgtg
     5461 cttcgatcaa tttgtctaca agctgagcga gcagatcttt gcccattaca agcaattggc
     5521 gggcagcatc ttcttggaca agcgattccg tttggagtgc gaggtactcg gtttcaactt
     5581 ccagtcgtat ccccgcaata atcgttatga gaccttgtta aaacaacgac atgtccaact
     5641 tttgggccgt tctattgact tgaacaaact gatcacacaa cgcataaacg ccaacatgca
     5701 caagagcatt gagttggcca tcagtcgttt cgagggcaac gatataaccg gcattgtgga
     5761 actcgaaggc ctgctagagg cgaatcgaat ttgccacaag ttgctaagca agtacttggc
     5821 cctggataac tttgacggca tggtaaagga ggctaaccac aatgtactgg cgccttacgg
     5881 cagaataaca ctgcacgtct ttgtggaatt gaactacgac tttctggtca attactgcta
     5941 taatgcagcc acgaatcgtt tcattcgaac caaagtgaat ctttcgtcgt cacaggctat
     6001 tcagcgtgag aagccgccac aaatgtccca ttactatctg tggggttcca agcagctgaa
     6061 tgctgcctat tccacacagt atggccagta cactggcttt gtaggttcac cgcatttcca
     6121 tgccatgtgc cgcctgctgg gctatcaggg catagcagtc gtcatggaca ttatactgaa
     6181 ggacattgtt aagccactga tccagggctc gctgctgcag ttcaccaaga cgctgatgat
     6241 tgccatgccc aagtcatgca aactaccccg ttgcgaatat ggttcgcctg gagtgctcag
     6301 ctactatcag gctcatctta cagacatcgt gcaatatccg gacgcgaaga cagagctatt
     6361 tcagtctttc cgggagtttg gcaacagcat tatcttttgc ctgctgatcg aacaggcgct
     6421 ctcgcaggag gaggtgtgcg atctactgca cgccgccctc ttccagaaca tctttcccag
     6481 acctttttgc aaagagaatg aaaaaccgga ggctaagcaa aagcgtcttg aagcacagtt
     6541 tgccaatctg cagatagttt caaacgtgga aaagattggc actgcaaagc aagcaatgat
     6601 agcacgagag ggagatttgc tgactcgtga gcgtctttgc tgtggcctca gcatttttga
     6661 agttattctc aaccgcgtga agagctattt ggacgaccct gtgtggtgtg gtcctccacc
     6721 ggccaacggg attatccacg tggatgagtg ctcggagttc catcgccttt ggtcggctct
     6781 gcagttcgtt tattgcattc cagtcagggg aacggagtac accatcgagg agctcttcgg
     6841 cgaaggtctg aactgggctg gctgcgtcat gatcgtactg ctgggacagc agcgtcgctt
     6901 cgaggctctg gacttttgct accacattct gcgggtacag cgcgtcgatg gcaaggatga
     6961 ggacgtaaag ggcattcaat tgaagcgaat ggtggatcgg attcgtcggt tccaagtatt
     7021 aaactcacaa atcttctcca ttctcaacaa gtatctaaag ggcggcgatg gcgagggctc
     7081 aaatgtggag cacgtgcggt gttttccacc gcctcagcat ccctcggtca tatcgtcctc
     7141 gtcgcactac caggatcccc agaagctgcg gcagagcata aacaattgag cggccgctcg
     7201 agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat
     7261 tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg
     7321 actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc agacatgata
     7381 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     7441 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     7501 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     7561 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc gaccgatgcc
     7621 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     7681 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg
     7741 ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc
     7801 ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg
     7861 tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt
     7921 gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg
     7981 cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca
     8041 gggacagctt caaggatcgc tcgcggctct taccagccag caaaaggcca ggaaccgtaa
     8101 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     8161 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     8221 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     8281 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     8341 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     8401 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8461 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8521 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     8581 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8641 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8701 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8761 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8821 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8881 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8941 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     9001 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     9061 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     9121 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     9181 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     9241 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     9301 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     9361 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     9421 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     9481 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     9541 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9601 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9661 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9721 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9781 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9841 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     9901 attaacctat aaaaataggc gtatcacgag gccctttcgt
//
