LOCUS       sequence-288450-        9062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmSmaug-A642H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9062)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 9062)
  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-DmSmaug-A642H-V5His6_C
FEATURES             Location/Qualifiers
     source          1..9062
                     /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             6347..6388
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6398..6415
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6438..6455)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6488..6622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6512..6531)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6566..6585
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7244..7832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7324..7343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8003..8794,8795..8863))
                     /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     8626..8645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8864..8968)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9036..9054
                     /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 ttgaattctg cagatatcat gaagtacgca actggaactg acaacgcgat
     3361 gacctccggc atcagcggcc aaaccaataa ctctaacagt gcatccaacg agatgcagcc
     3421 aaccacatca acgcctactg cagcccacaa ggaggccaca tcaacggcca cgacgacggc
     3481 gacatacgcc aatggcaatc ccaattctaa cgccaatccc agccagagcc agccgtccaa
     3541 tgcgctgttt tgcgagcagg tgaccacagt gacaaatctc ttcgagaagt ggaacgactg
     3601 cgagcgaacc gtcgtgatgt acgcgctgct caagcggctg cgctacccga gcctcaagtt
     3661 tctacagtat tctatcgaca gcaacctgac ccagaacctg ggcacctcgc agaccaatct
     3721 gagcagcgtg gttattgaca tcaatgccaa caatccggtg tacctgcaga atctactcaa
     3781 cgcctacaag acgttccagc catgtgacct gctcgacgcc atgtcctcct cgtcgtccga
     3841 caaggactcg atgccatgtt acggcagtga cttccagatc accacatcag cgcagtgtga
     3901 cgagcgaaag ctgtatgccc gcaaggagga cattctgcat gaagtactga acatgctgcc
     3961 tctgctgaag ccgggcaatg aggaggccaa gcttatctac ctgaccctca taccagttgc
     4021 cgtcaaggac accatgcagc aaattgtgcc cacggagttg gtgcagcaga tcttctcgta
     4081 cctactcatc catccagcta tcaccagcga ggacagacgt tcgctcaaca tttggctgcg
     4141 tcacttggag gatcatatcc aagcggctgc ggcgggcctg acaaatcgca gttacttcct
     4201 gcagccctcg ccgcaactgg tcgctggtgg tagctcaaca ggcagtggta gctgttcctc
     4261 ttcggcgacc agctcttcga cagcatcctg ttcgtcggtg gcctcatcct cgttgtgccc
     4321 cgcaagcggc agccgttcct cacgcaccaa cgactggcaa acgatcgccc cgcccagcaa
     4381 gcaattgcaa aacaaattgg ccggtgattg gcgtggcaat ggaggcggca gctccagcgg
     4441 gtcgatcaat ccactctgtg ataatcttaa tggtattacc ctgaacgaat tagcttctag
     4501 tcagaacagt ctcggcctgt cgctggaggg cagctcttcc ctggtcaacg gtgtggtcgc
     4561 tggtgcaggc tctatgttgg gaatcgctgg tggcgacgat catgacacgt catttagcaa
     4621 gaatggcacg gagattctcg attttgaccc ggtgacggcg gacatgggtg aggcctgtag
     4681 tctggccagc agtagccttt gtggacgtaa cggtggaaat cctgttgagg atcgatcgca
     4741 acctccgccg aatcttcagc agcaattgct tcagcctcct ccctatgcct cgatccttat
     4801 gggaaacgtg ggcgatcagt ttggagagat caaccggtgg agcctggaca gcaagattgc
     4861 agccctgaag acgcggcgct ctaacagcct caccacgcaa acgatctcta gctgttccag
     4921 ctcgtctaac tcatcggtga tcacggtcaa tgacaactgc tccaattcaa cggagaatct
     4981 ggcacagttt gccaacaagc cccgtagttt ttccttatcg attgagcatc agcgcggcgc
     5041 cttgatgaac agcggttccg atactcgtct ggatgaattc aagcccaatt atattaagtt
     5101 ccacacgcgc aacgtgggca tgtcgggaat cggactctgg ctaaaatcct tgcgtctgca
     5161 caagtacatc gagctgttca agaacatgac gtacgaggaa atgctcttga tcaccgagga
     5221 ctttctgcag agtgttggcg tcaccaaggg ccactcccac aagcttgccc tttgcattga
     5281 taagctgaag gaacgggcca atattcttaa ccgggtagaa caagaactgc tatcgggtca
     5341 aatggagctg agcacggcgg ttgaggagct aacaaatatt gtgctgacac caatgaagcc
     5401 actggagtct ccaggaccac cggaggaaaa cattggtctg cgttttctta aggtcattga
     5461 cattgtgacc aatacgttgc agcaggatcc gtatgctgtc caagacgacg agactttggg
     5521 tgtgcttatg tggattctag accgttctat ccacaacgag gcatttatga atcatgccag
     5581 tcaactcaag gacctgaagt ttaagttgtc caagatgaag atctccatgg ttcccaaaat
     5641 gcatcacgtt aagccagccg gcgtcgggcc aaacaatggg aacattaaca agccaagatg
     5701 gaatggtaag actcgcaaat gcgatactaa aaatggcagc aacgatagga ttaacaaccg
     5761 caaaaactcc aacgacatgc taaacttttc gctaaattgc ctgccacacc cgctgcccca
     5821 tcattcgcag caggctccgc cgcctctgcc acagtttgac tacaacggct atggcggcgg
     5881 tccatcccat cagccgcagt acaagagctc ctcgtaccca agcttcatgg gtaatcctca
     5941 gcagcagcca ccaccgccgc cgtctagtaa gtcgcatcat catccccagc agatgcaaca
     6001 gatgctgcag caacacaacc actttccagc gctgcctcag cagacgccac cacaatcgca
     6061 tcgccggtcg ctgaataatc ttattctggt ggccggaggt ccgcagcaac cgcagcaatt
     6121 gatctttaag cccggtcagg gagtgctcac caacaacgga tctaacgaca acctggtatt
     6181 ggaacgcaat cagcagtcgc agcagcagag aaaacttagc ggaggagtgt ccagtgcgga
     6241 gcaacaacca aagaagacca tggccgctgt tgtgatggaa aatttggcca aatttgatca
     6301 acattttacg ctattccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc
     6361 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     6421 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     6481 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     6541 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     6601 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     6661 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     6721 ttatgatcag tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     6781 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     6841 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga
     6901 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg
     6961 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg
     7021 ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca
     7081 ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca
     7141 ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc
     7201 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7261 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7321 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7381 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     7441 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7501 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7561 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7621 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7681 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7741 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7801 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7861 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7921 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7981 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8041 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8101 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8161 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8221 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8281 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     8341 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8401 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8461 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8521 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8581 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     8641 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8701 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8761 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8821 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8881 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8941 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     9001 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     9061 gt
//
