LOCUS       sequence-289514-        7704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-12xmut_1-539.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7704)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7704)
  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-DmGW182-12xmut_1-539_O
FEATURES             Location/Qualifiers
     source          1..7704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..2523
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2543..2545,2546..2548,2549..3259)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2543..2545,2546..2548,2549..3259)
                     /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(2588..2609)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2849..2868)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3196..3217
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4944..4985
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4995..5012
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5035..5052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5085..5219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5109..5128)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5163..5182
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5841..6429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5921..5940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6600..7391,7392..7460))
                     /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     7223..7242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7461..7565)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7633..7651
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taaaaaaaat catgaatggc atcaactctg aatcaaatct ttgcagatgc acctacttct
       61 catttccact gtcacatcat ttttccagat ctcgctgcct gttatgtggc ccacaaacca
      121 agacacgttt tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacatatcaa
      181 tatgtacatt cgagaaagaa gcgatcaaag aagcgtcttc gggcgagtag gagaatgcgg
      241 aggagaagga gaacgagctg atctagtatc tctccacaat ccaatgccaa ctgaccaact
      301 ggccatattc ggagcaattt gaagccaatt tccatcgcct ggcgatcgct ccattcttgg
      361 ctatatgttt ttcaccgttc ccggggccat tttcaaagac tcgtcggtaa gataagattg
      421 tgtcactcgc tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg
      481 gcgagtattt tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt
      541 tctacaaaaa catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag
      601 attgtatgta tataccttta atgtattctc attttcttat gtatttataa tggcaatgat
      661 gatactgatg atattttaag atgatgccag accacaggct gatttctgcg tcttttgccg
      721 aacgcagtgc atgtgcggtt gttgtttttt ggaatagttt caattttcgg actgtccgct
      781 ttgatttcag tttcttggct tattcaaaaa gcaaagtaaa gccaaaaaag cgagatggca
      841 ataccaaatg cggcaaaacg gtagtggaag gaaaggggtg cggggcagcg gaaggaaggg
      901 tggggcgggg cgtggcgggg tctgtggctg ggcgcgacgt caccgacgtt ggagccactc
      961 ctttgaccat gtgtgcgtgt gtgtattatt cgtgtctcgc cactcgccgg ttgttttttt
     1021 ctttttatct cgctctctct agcgccatct cgtacgcatg ctcaacgcac cgcatgttgc
     1081 cgtgtccttt atgcgtcatt ttggctcgaa ataggcaatt atttaaacaa agattagtca
     1141 acgaaaacgc taaaataaat aagtctacaa tatggttact tattgccatg tgtgtgcagc
     1201 caacgatagc aacaaaagca acaacacagt ggctttccct ctttcacttt ttgtttgcaa
     1261 gcgcgtgcga gcaagacggc acgaccggca aacgcaatta cgctgacaaa gagcagacga
     1321 agttttggcc gaaaaacatc aaggcgcctg atacgaatgc atttgcaata acaattgcga
     1381 tatttaatat tgtttatgaa gctgtttgac ttcaaaacac acaaaaaaaa aaataaaaca
     1441 aattatttga aagagaatta ggaatcggac agcttatcgt tacgggctaa cagcacaccg
     1501 agacgaaata gcttacctga cgtcacagcc tctggaagaa ctgccgccaa gcagagagag
     1561 agagaaaaag agggagagca gcttagaccg catgtgcttg tgtgtgaggc gtctctctct
     1621 tcgtctcctg tttgcgcaaa cgcatagact gcactgagaa aatcgattac ctatttttta
     1681 tgaatgaata tttgcactat tactattcaa aactattaag atagcaatca cattcaatag
     1741 ccaaatacta taccacctga gcgatgcaac gaaatgatca atttgagcaa aaatgctgca
     1801 tatttaggac ggcatcatta tagaaatgct tcttgctgtg tacttttctc tcgtctggca
     1861 gctgtttcgc cgttattgtt aaaaccggct taagttaggt gtgttttcta cgactagtga
     1921 tgcccctact agaagatgtg tgttgcacaa atgtccctga ataaccaatt tgaagtgcag
     1981 atagcagtaa acgtaagcta atatgaatat tatttaactg taatgtttta atatcgctgg
     2041 acattactaa taaacccact ataaacacat gtacatatgt atgttttggc atacaatgag
     2101 tagttgggga aaaaatgtgt aaaagcaccg tgaccatcac agcataaaga taaccagctg
     2161 aagtatcgaa tatgagtaac ccccaaattg aatcacatgc cgcaactgat aggacccatg
     2221 gaagtacact cttcatggcg atatacaaga cacacacaag cacgaacacc cagttgcgga
     2281 ggaaattctc cgtaaatgaa aacccaatcg gcgaacaatt catacccata tatggtaaaa
     2341 gttttgaacg cgacttgaga gcggagagca ttgcggctga taaggtttta gcgctaagcg
     2401 ggctttataa aacgggctgc gggaccagtt ttcatatcac taccgtttga gttcttgtgc
     2461 tgtgtggata ctcctcccga cacaaagccg ctccatcagc cagcagtcgt ctaatccaga
     2521 gaccccggat cggggtacca acatggtgag caagggcgag gagctgttca ccggggtggt
     2581 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2641 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2701 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2761 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2821 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2881 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2941 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3001 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3061 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3121 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3181 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3241 catggacgag ctgtacaagc caagcttgaa ttctgcagat atccagcaca gtggcggccg
     3301 catgcgtgaa gccctttttt cccaagatgc cgcggcctgt cagcatgtta accaggatac
     3361 taattgggaa gttcccagtt cgccagaacc agccaataag gatgcacccg gtccaccaat
     3421 gtggaagcca agcattaaca atggtactga tctttgggag tccaatttga gaaacggagg
     3481 tcagccggcc gcacagcaag ttccaaagcc gtcggcggct catacaccat cctctaactt
     3541 aggtggaaca gcagctgagg acgacgatgg cgccgatagt agtagtgtgt ggactggagg
     3601 agctgttagc aacgcgggat ccggagctgc agtgggagta aaccaagccg gagttaatgt
     3661 cggtccaggc ggtgttgttt cgtctggcgg acctcaggcg gcacaaggtg tcgttggcgt
     3721 cggacttgga tcaactggag gtaacgggtc aagcaatata actggatcgt ctggagtcgc
     3781 aacaggtagt agcggaaact ccagcaacgc tggtaacgct gcggcagacc ctcgtgaaat
     3841 acgccctttg ggagttggtg gctccatgga tattcgaaat gttgaacatc gcggcggtaa
     3901 cggttctgga gcaacttcga gcgatccacg agacattcgc atgatcgatc cgcgtgaccc
     3961 tattcgagga gatccccgtg gaatatctgg tcgtcttaat gggacctctg aaatggccgc
     4021 tcatcatcca caaatgtccc ataaccagtt gcaaggtatc aacaaaatgg ttggtcaaag
     4081 tgtagcaact gccagcacca gtgtcggaac atctggctcg ggcatcggtc ctggaggtcc
     4141 cggtcctagt acagtatcag gcaatatccc aacacaggcg gcgcctgctc aaccggtaag
     4201 cgttggtgta agtggtccca aagacatgtc aaaacagata agtgcagcgg aggaaccatc
     4261 accaccgcct cagcgtcgca gtattcctaa ctacgatgat ggtacatcgt tggcggctca
     4321 gcaaactcgt gttcccgctg caagcggtca ctggaaagac atgactgatt cgataggtcg
     4381 tagtagtcat ctcatgcgtg gccaaagcca aacgggaggt ataggaatag ccggcgttgg
     4441 aaatagcaat gttccagtgg gagccaatcc aagtaatcct ataagcagtg tagttggacc
     4501 tcaagcccgg attccatctg tgggcggcgt acaacacaaa ccagacggcg gcgctatgtg
     4561 ggtgcattcc ggcaatgtag gtggcagaaa taatgttgct gctgttacta ctgcagcgga
     4621 tgacactcat agcgttaatg tcggcgctcc cagcagtggc agtgtatcca gcaacaattg
     4681 ggttgatgac aagtccaact caaccttggc acaaaactct tggagcgacc cggcccctgt
     4741 tggagttagt gccgcgaata agcaaagcaa accgccaagc aatagtgctt catcagctgc
     4801 gagcactgct gcgggcgtgg tggatggggt tgatctagga tctgagtgga acacgcacgg
     4861 ggggattatt ggaaaatctc agcaacaaca aaaactagcg ggacttaacg tgggaatgta
     4921 atctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     4981 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     5041 ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa
     5101 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     5161 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     5221 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     5281 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg accgatgccc
     5341 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     5401 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     5461 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     5521 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     5581 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
     5641 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     5701 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     5761 ggacagcttc aaggatcgct cgcggctctt accagccagc aaaaggccag gaaccgtaaa
     5821 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5881 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5941 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6001 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     6061 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6241 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6361 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6421 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6481 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6541 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6601 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6661 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6721 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6781 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6841 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6901 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6961 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7021 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7081 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7141 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7201 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7261 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7321 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7381 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7441 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7501 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7561 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7621 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaaggaat taattctata
     7681 ttctaaaaac acaaatgata cttc
//
