LOCUS       sequence-288363-        9178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmBrat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9178)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 9178)
  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"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmBrat_B
FEATURES             Location/Qualifiers
     source          1..9178
                     /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     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6463..6504
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6514..6531
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6554..6571)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6604..6738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6628..6647)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6682..6701
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7360..7948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7440..7459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8119..8910,8911..8979))
                     /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     8742..8761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8980..9084)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9152..9170
                     /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 ttaatggcgt cctcaccgac accatctctg gactcgatgc ggggcggggc
     3361 gaactcgatt gaatcgtacg aacacgccgg ctatctatca gactctccgc tcacgttgag
     3421 cggttcatcg ccaccggcaa gcgattcggc aatctgcagt gatgagtaca cgggcggcag
     3481 cagcgttaaa tcacgctctg aggtgactgt aatcaatggc catcatccca tttcggcctc
     3541 cgtatcgtcc agctcttcgg ccagctcttc atcctgctcg tcgtcatcgt cctcatcctc
     3601 gtcttcgtca tcgtccagct cctcgaccag cggcctcagc ggttgcggca gtaccagcag
     3661 cagtgtgatc agcgccaaca acgttgcgag cagcaatgga ccaggcgtta tcggcagcaa
     3721 tcttcagtcc tcaaacaatg gcggcaactc gggcatcagc agtctggtcg tgggtgcggg
     3781 caagggaagc aacagcagca gcaacagctc ctcgagcaac acatccgcca atggatctcc
     3841 gccacgttgc accgcctgca agagcaagtg cagcgatgcg gtggccaagt gcttcgaatg
     3901 ccagagctac ctgtgcgcca attgtgtaac tgctcatgag ttcatgcact gcttcaacgg
     3961 acacaatgtg tgcctgatta agggttttga ggcgagcacc actggcactc cgctgtcggt
     4021 tggatcgcct agcaacaatc cggcgtcgaa tgagttcaag tacgccagtt cgctgaccat
     4081 gatgctgcag cagcagcaac agctcgactc gcaacaacag caacagcagc aacaattgct
     4141 gccggcgcag cctatgtcgc aactctcgaa gattgtactg gcagccgccg cgcaggcaaa
     4201 ttcccaggag caacagcgag aggatagcat ctatggttca ctgcacccac aacaacaaca
     4261 acagcagcag caacagcagc agcgacaact cttctgcccc cgacacaaac aggagctgct
     4321 caagttcagt tgccgcacct gctgcatcct ggtgtgcaag gagtgcattg tactggagca
     4381 ctccacaggt ctccacgaac tggagaacgt gcagtccccc ggaatgacaa cctctacggg
     4441 atccacagcc aacgagtcag ctttgcagac tctgctcgcc gatatgcgtg gcaaaattgg
     4501 cgagattgtc ggcattgctg gaaactcgga ccagaatctc accaaggtga agctgcagta
     4561 ccaaaaggct cacaacgagc tgaacgagac gcaccagttc ttcgcttcca tgctggacga
     4621 gcgcaagacc gaattgctca aggaactgga gacgctgtac acggccaagg ttaacagcaa
     4681 caattcgtgg cagcaacgct cccgcgactt gatcgacaag ggactggcca cctgcgaggc
     4741 ggtcgagcgg agtcctgctc cgccatcatc cttgctgaca gaggcccttc tcctgcgcaa
     4801 atccctggag caacaactac agacgggcat tcaggagatg cagctaccat tcgagatcga
     4861 atttatgtcc aactatcagt cgattcaggc cggcgtgcgc aacacctttg gttacattcg
     4921 ggcaaacagc tcagatggtg gacctactgg catgagcctg acgagcaatg gccacggaaa
     4981 gcagccgcca attgcccgac caacacagag cgccagcaac agcagtgcga gcagcgcggg
     5041 cagtggtcac catggccacc accaacagtc acaccatcat ggccaccaca atcatcacca
     5101 gacggcccat catcaacagc tgcaggcgca atcgtcgctg catggcttgg gtcttggatt
     5161 gagtggcgcc agtttgttgg atagcagctc tagcgcaggc ggagcagtgg gagccttcag
     5221 caacggagga cttctgctgg gcggtcgcga tcggaacgct ctggcagtgg aacagcattt
     5281 cggtgagctg atgcccaagc gaggaggagg tggctacacg ggcagcaatg gatcggccac
     5341 cagtgccgtg gcccactaca atccctacga gaagtggagc aacggcggta gcgacaatct
     5401 gttttcatcg gtgacaagcg gcgtgtccgg tagctctgca gtggccgatg catttgccag
     5461 cctgtctgct gttggaggat ccgtggttag tggcgctgga gctggcggaa gtactgtgag
     5521 ctcagagtcg ttgctggacc tcaccaacaa gctgctttcg gccactatct atccgcccaa
     5581 gtcgcagatc aagcgacaga agatgattta ccactgcaag tttggcgaat tcggcgtgat
     5641 ggagggccag ttcacggagc ccagcggtgt ggcggtaaat gcccagaatg acatcattgt
     5701 tgcggatacg aacaaccatc gcattcagat cttcgacaag gagggacgct tcaagttcca
     5761 gtttggcgag tgcggcaagc gtgactcgca gctgctctat ccgaaccgcg tggccgtggt
     5821 gcgcaattcc ggcgatatta ttgtcaccga gcgctcgccc acacaccaga tacagatcta
     5881 caatcagtac ggccagtttg tgaggaaatt cggggccacc attctgcagc atcctcgcgg
     5941 cgtgaccgtg gacaacaagg gacggatcat tgtggtggaa tgcaaggtga tgcgtgtgat
     6001 catctttgat cagaacggca atgtgctgca caagttcggg tgctctaagc acctcgagtt
     6061 ccccaacggc gtggtggtca acgacaagca ggagatcttc atcagcgaca atcgggcgca
     6121 ctgtgtcaag gtgttcaact acgagggcca gtatctgcgg cagattggcg gcgagggtat
     6181 taccaactac ccaattggcg tcgggatcaa ctcgaacggg gagatcctca tcgcggacaa
     6241 ccacaacaac ttcaacctga cgatcttcac gcaggacggg cagctgatct cggcgctcga
     6301 gtcgaaggtg aagcacgcgc agtgcttcga tgtggcgctc atggacgacg gcagtgtggt
     6361 gctggccagc aaggactatc ggctctacat atatcgctat gtccaactgg cgccagtggg
     6421 tatgtaagcg gccgctcgag tctagagggc ccgcggttcg aaggtaagcc tatccctaac
     6481 cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca ttgagtttaa
     6541 acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc agcctcgatc
     6601 gaggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     6661 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     6721 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     6781 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     6841 gatcagtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     6901 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     6961 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat
     7021 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac
     7081 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga
     7141 cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat
     7201 gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca
     7261 ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta ccagccagca
     7321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     7561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7801 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7921 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7981 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     8041 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8101 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8161 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8221 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8281 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8341 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8401 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
     8461 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8521 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8581 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8641 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8701 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     8761 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8821 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8881 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8941 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     9001 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9061 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9121 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgt
//
