LOCUS       sequence-333036-        6946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmBelle_1-298.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6946)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6946)
  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-DmBelle_1-298_B
FEATURES             Location/Qualifiers
     source          1..6946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /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(2630..2651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2891..2910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3238..3259
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4234..4275
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4285..4302
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4325..4342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4375..4509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4399..4418)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4453..4472
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5128..5716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5208..5227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5887..6678,6679..6747))
                     /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     6510..6529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6748..6852)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6920..6938
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 catcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2641 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2701 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2761 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2821 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2881 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2941 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3001 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3061 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3121 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3181 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3241 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3301 gccaagcttg atgagtaatg ctattaacca aaatggcaca ggtctagagc agcaagttgc
     3361 tggtctggac ttgaatggcg gctctgctga ctacagcggc cccataacca gcaagacttc
     3421 cactaactcg gtcaccggtg gtgtgtatgt gcccccgcac cttcgtggtg gtggtggcaa
     3481 taacaacgca gcggacgcag agagccaggg ccaaggacag ggccagggtc agggcttcga
     3541 ctccagatcc gggaatccgc gccaggagac tagggaccct caacagtcgc gcggaggcgg
     3601 aggcgaatac cgcagaggcg gcggcggtgg tggtcgcggc ttcaatcgcc agagcggcga
     3661 ctacggttac ggaagcggcg gcggaggacg acgtggaggc ggtggacgtt tcgaggacaa
     3721 ctacaacggt ggtgaattcg attcgcgtcg cggcggtgac tggaatcgca gcggcggagg
     3781 aggtcgagga tttggacgcg gaccatcgta ccgcggcggt ggcggcggaa gcggtagcaa
     3841 cctcaacgag cagactgccg aggatggcca ggcacagcag cagcagcagc cgcgcaacga
     3901 tcgttggcag gagccagaac gtcctgccgg attcgatggc agcgagggcg gacagtctgc
     3961 cggcggcaac agaagctaca acaatcgcgg cgagcgcgga ggcggcggtt acaacagccg
     4021 ctggaaggag ggcggcggct caaacgtcga ctacacaaag ctgggcgccc gagatgagcg
     4081 cctggaggtg gagctgttcg gcgtgggcaa tacgggaatc aattttgaca aatacgagga
     4141 tatacccgtg gaggcgacgg gccagaatgt gcccccgaac atcacatcgt tcgattaggc
     4201 ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc
     4261 ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga
     4321 tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca
     4381 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4441 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4501 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     4561 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg
     4621 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     4681 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     4741 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
     4801 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
     4861 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg
     4921 ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct
     4981 cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga
     5041 cgaccatcag ggagatgacg accatcaggg acagcttcaa ggccagcaaa aggccaggaa
     5101 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5161 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5221 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5281 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5341 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5401 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5461 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5521 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5581 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5641 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5701 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5761 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5821 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5881 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5941 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6001 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6061 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6121 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6181 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6241 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6301 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6361 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6421 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6481 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     6541 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6601 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6661 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6721 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6781 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6841 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6901 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgt
//
