LOCUS       sequence-333046-        8336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmSmaug_1-762.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8336)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8336)
  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_1-762-V5His6_C
FEATURES             Location/Qualifiers
     source          1..8336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     86..104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        168..2677
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2697..2699,2700..2702,2703..3413)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2697..2699,2700..2702,2703..3413)
                     /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(2742..2763)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3003..3022)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3350..3371
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5754..5795
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5805..5822
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5845..5862)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5895..6029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5919..5938)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5973..5992
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6630..7218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6710..6729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7389..8180,8181..8249))
                     /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     8012..8031
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8250..8336,1..18))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
       61 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
      121 attaattcta tattctaaaa acacaaatga tacttctaaa aaaaaatcat gaatggcatc
      181 aactctgaat caaatctttg cagatgcacc tacttctcat ttccactgtc acatcatttt
      241 tccagatctc gctgcctgtt atgtggccca caaaccaaga cacgttttat ggccattaaa
      301 gctggctgat cgtcgccaaa caccaaatac ataatgaata tgtacacatt cgagaaagaa
      361 gcgatcaaag aagcgtcttc gggcggagta ggagaatgcg gaggagaagg agaacgagct
      421 gatctagtat ctctccacaa tccaatgcca actgaccaac tggccatatt cggagcaatt
      481 tgaagccaat ttccatcgcc tggcgatcgc tccattcttg gctatatgtt tttcaccgtt
      541 acccggggcc attttcaaag actcgtcggc aagataagat tgtgtcactc gctgtctctc
      601 ttcatttgtc gaagaatgct gaggaatttc gcgatgacgt cggcgagtat tttgaagaat
      661 gagaataatt tgtatttata cgaaaatcag ttagtggaat tttctacaaa aacatgttat
      721 ctatagataa ttttgttgca aaatatgttg actatgacaa agattgtatg tatatacctt
      781 taatgtattc tcattttctt atgtatttat aatggcaatg atgatactga tgatatttta
      841 agatgatgcc agaccaaaag gcttgaattt ctgcgtcttt tgccgaacgc agtgcatgtg
      901 caattgttgt tttttggaat attcaatttt cggactgtcc gctttgattt cagtttcttg
      961 gcttattcaa aaagcaaagt aaagccaaaa aagcgagatg gcaataccaa atgcggcaaa
     1021 acggtagtgg aaggaaaggg gtgcggggca gcggaaggaa gggtggggcg gggcgtggcg
     1081 gggtctgtgg ctgggcgcga cgtcaccgac gttggagcca ctcctttgac catgtgtgcg
     1141 tgtgtgtatt attcgtgtct cgccactcgc cggttgtttt tttcttttta tgctgcgctc
     1201 tctctagcgc catctcgctt acgcatgctc aacgcaccgc atgttgccgt ttccttttat
     1261 gcgtcatttt ggctcgaaat aggcaattat ttaaacaaag attagtcaac gaaaacgcta
     1321 aaataaataa gtctacaata tggttactta ttgccatgtg tgtgcagcca acgatagcaa
     1381 caaaagcaac aacacaggtg gctttccctc tttcactttt tgtttgcaag ccgcgtgcga
     1441 gcaagacggc acgaccggca aacgcaatta cgctgacaaa gagcagacga agttttggcg
     1501 aaaaacatca aggcgcctga tacgaatgca tttgcaataa caattgcgat atttaatatt
     1561 gtttatgaag ctgtttgact tcaaaacaca caaaaaaaaa aataaaacaa attatttgaa
     1621 agagaattag gaatcggacg cttatcgtta gggtaacaac aagaaatgct tactgagtca
     1681 cagcctctgg aaaactgccg caagccagag agagagagaa aaagagggag agcagcttag
     1741 accgcatgtg cttgtgtgtg aggcgtctct ctcttcgtct ctgttgcgca aacgcataga
     1801 ctgcactgaa aaaatcgatt acctattttt tatgaatgaa tatttgcact attactattc
     1861 aaaactatta agatagcaat cacattcaat agccaaatac tataccacct gagcgatgca
     1921 acgaaatgat caatttgagc aaaaatgctg catatttagg acggcatcat tatagaaatg
     1981 cttcttgctg tgtacttttc tctcgtctgg cagctgtttc gccgttattg ttaaaaccgg
     2041 cttaagttag gtgtgttttc tacgactagt gaatgcccta ctagaagatg tgtgttgcac
     2101 aaaatgtccc tggaataacc aatttgaagt gcagatagca gtaaacgtaa gctaatatga
     2161 atattattta actgtaatgt tttaatatcg ctggacatta ctaataaacc cactataaac
     2221 acatgtacat atgtatgttt tggcatacaa tgagtagttg gggaaaaaat gtgtaaaagc
     2281 accgtgacca tcacagcata aagataacca gctgaagtat cgaatatgag taacccccaa
     2341 attgaatcac atgccgcaac tgataggacc catggaagta cactcttcat ggcgatatac
     2401 aagacacaca caagcacgaa cacccagttg cggaggaaat tctccgtaaa tgaaaaccca
     2461 atcggcgaac aattcatacc catatatggt aaaagttttg aacgcgactt gagagcggag
     2521 agcattgcgg ctgataaggt tttagcgcta agcgggcttt ataaaacggg ctgcgggacc
     2581 agttttcata tcactaccgt ttgagttctt gtgctgtgtg gatactcctc ccgacacaaa
     2641 gccgctccat cagccagcag tcgtctaatc cagagacccc ggatcggggt accatcatgg
     2701 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2761 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2821 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2881 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2941 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3001 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3061 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3121 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3181 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3241 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3301 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3361 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagccaagct
     3421 tgaattctgc agatatcatg aagtacgcaa ctggaactga caacgcgatg acctccggca
     3481 tcagcggcca aaccaataac tctaacagtg catccaacga gatgcagcca accacatcaa
     3541 cgcctactgc agcccacaag gaggccacat caacggccac gacgacggcg acatacgcca
     3601 atggcaatcc caattctaac gccaatccca gccagagcca gccgtccaat gcgctgtttt
     3661 gcgagcaggt gaccacagtg acaaatctct tcgagaagtg gaacgactgc gagcgaaccg
     3721 tcgtgatgta cgcgctgctc aagcggctgc gctacccgag cctcaagttt ctacagtatt
     3781 ctatcgacag caacctgacc cagaacctgg gcacctcgca gaccaatctg agcagcgtgg
     3841 ttattgacat caatgccaac aatccggtgt acctgcagaa tctactcaac gcctacaaga
     3901 cgttccagcc atgtgacctg ctcgacgcca tgtcctcctc gtcgtccgac aaggactcga
     3961 tgccatgtta cggcagtgac ttccagatca ccacatcagc gcagtgtgac gagcgaaagc
     4021 tgtatgcccg caaggaggac attctgcatg aagtactgaa catgctgcct ctgctgaagc
     4081 cgggcaatga ggaggccaag cttatctacc tgaccctcat accagttgcc gtcaaggaca
     4141 ccatgcagca aattgtgccc acggagttgg tgcagcagat cttctcgtac ctactcatcc
     4201 atccagctat caccagcgag gacagacgtt cgctcaacat ttggctgcgt cacttggagg
     4261 atcatatcca agcggctgcg gcgggcctga caaatcgcag ttacttcctg cagccctcgc
     4321 cgcaactggt cgctggtggt agctcaacag gcagtggtag ctgttcctct tcggcgacca
     4381 gctcttcgac agcatcctgt tcgtcggtgg cctcatcctc gttgtgcccc gcaagcggca
     4441 gccgttcctc acgcaccaac gactggcaaa cgatcgcccc gcccagcaag caattgcaaa
     4501 acaaattggc cggtgattgg cgtggcaatg gaggcggcag ctccagcggg tcgatcaatc
     4561 cactctgtga taatcttaat ggtattaccc tgaacgaatt agcttctagt cagaacagtc
     4621 tcggcctgtc gctggagggc agctcttccc tggtcaacgg tgtggtcgct ggtgcaggct
     4681 ctatgttggg aatcgctggt ggcgacgatc atgacacgtc atttagcaag aatggcacgg
     4741 agattctcga ttttgacccg gtgacggcgg acatgggtga ggcctgtagt ctggccagca
     4801 gtagcctttg tggacgtaac ggtggaaatc ctgttgagga tcgatcgcaa cctccgccga
     4861 atcttcagca gcaattgctt cagcctcctc cctatgcctc gatccttatg ggaaacgtgg
     4921 gcgatcagtt tggagagatc aaccggtgga gcctggacag caagattgca gccctgaaga
     4981 cgcggcgctc taacagcctc accacgcaaa cgatctctag ctgttccagc tcgtctaact
     5041 catcggtgat cacggtcaat gacaactgct ccaattcaac ggagaatctg gcacagtttg
     5101 ccaacaagcc ccgtagtttt tccttatcga ttgagcatca gcgcggcgcc ttgatgaaca
     5161 gcggttccga tactcgtctg gatgaattca agcccaatta tattaagttc cacacgcgca
     5221 acgtgggcat gtcgggaatc ggactctggc taaaatcctt gcgtctgcac aagtacatcg
     5281 agctgttcaa gaacatgacg tacgaggaaa tgctcttgat caccgaggac tttctgcaga
     5341 gtgttggcgt caccaagggc gcctcccaca agcttgccct ttgcattgat aagctgaagg
     5401 aacgggccaa tattcttaac cgggtagaac aagaactgct atcgggtcaa atggagctga
     5461 gcacggcggt tgaggagcta acaaatattg tgctgacacc aatgaagcca ctggagtctc
     5521 caggaccacc ggaggaaaac attggtctgc gttttcttaa ggtcattgac attgtgacca
     5581 atacgttgca gcaggatccg tatgctgtcc aagacgacga gactttgggt gtgcttatgt
     5641 ggattctaga ccgttctatc cacaacgagg catttatgaa tcatgccagt caactcaagg
     5701 acctgaagtt taagttgtcc aaacgctcga gtctagaggg cccgcggttc gaaggtaagc
     5761 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     5821 attgagttta aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat
     5881 cagcctcgat cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     5941 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     6001 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     6061 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     6121 tggctgatta tgatcagtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     6181 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     6241 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     6301 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     6361 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     6421 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     6481 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     6541 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggccagca aaaggccagg
     6601 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6661 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6721 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6781 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6841 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6901 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6961 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7021 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7081 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7141 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7201 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7261 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7321 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7381 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7441 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7501 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7561 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7621 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7681 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     7741 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7801 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7861 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7921 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7981 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     8041 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8101 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8161 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8221 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8281 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataa
//
