LOCUS       sequence-141584-        5190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  targeting vector for C-terminal eGFP tag on Drosophila melanogaster 
            Act5C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5190)
  AUTHORS   Kunzelmann S, Bottcher R, Schmidts I, Forstemann K
  TITLE     A Comprehensive Toolbox for Genome Editing in Cultured Drosophila 
            melanogaster Cells.
  JOURNAL   G3 (Bethesda). 2016 Jun 1;6(6):1777-85. doi: 10.1534/g3.116.028241.
  PUBMED    27172193
REFERENCE   2  (bases 1 to 5190)
  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"; doi: "10.1534/g3.116"; 
            journalName: "G3 (Bethesda)"; date: "2016-06-1- 1"; volume: "6"; 
            issue: "6"; pages: "1777-85"
COMMENT     Sequence Label: pIS1
FEATURES             Location/Qualifiers
     source          1..5190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(857..859,860..862,863..1576)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(857..859,860..862,863..1573)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(902..923)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1163..1182)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1510..1531
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1583..1616)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             1893..2291
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(2340..2373)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        complement(join(3022..3028,3029..3046,3047..3052))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3067..3088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3208..3225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3379..3967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3459..3478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4138..4929,4930..4998))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4761..4780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4999..5103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5171..5189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgctccgt gacatcaagg agaagctgtg
      361 ctatgttgcc ctcgactttg agcaggagat ggccaccgct gccagcagct cctcgttgga
      421 gaagtcctac gagctgcccg acggacaggt gatcaccatc ggcaacgagc gtttccgctg
      481 ccccgaggcc ctgttccagc cctcgttctt gggaatggag gcttgcggca tccacgagac
      541 cacctacaac tccatcatga agtgtgatgt ggatatccgt aaggatctgt atgccaacac
      601 cgtgctgtcc ggtggcacca ccatgtaccc tggcatcgcc gaccgtatgc agaaggagat
      661 caccgccctg gcaccgtcga ccatgaagat caagatcatt gccccgccag agcgcaagta
      721 ctctgtctgg atcggtggct ccatcctggc ttcgctgtcc accttccagc agatgtggat
      781 ctccaagcag gagtacgacg agtccggccc ctccattgtg caccgcaagt gcttcggatc
      841 ttccggatgg ctcgagatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      901 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      961 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1021 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1081 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1141 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1201 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1261 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1321 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1381 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1441 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1501 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1561 cgagctgtac aagtaaacta gtgaagttcc tatactttct agagaatagg aacttcaaat
     1621 aaacatatgc tgttggaata tactattcaa cctacaaaaa taacgttaaa caacactact
     1681 ttatatttga tatgaatggc cacacctttt atgccataaa acatattgta agagaatacc
     1741 actcttttta ttccttcttt ccttcttgta cgttttttgc tgtgagtagg tcgtggtgct
     1801 ggtgttgcag ttgaaataac ttaaaatata aatcataaaa ctcaaacata aacttgacta
     1861 tttatttatt tattaagaaa ggaggtaccg gtatggccaa gcctttgtct caagaagaat
     1921 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     1981 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     2041 attttactgg gggaccttgc gcagaactcg tggtgctggg cactgctgct gctgcggcag
     2101 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     2161 gcggacggtg ccgacaggtt cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     2221 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     2281 gggagggcta gaaattatta aaaataaaac tttaaaaata aatttgtttc taacatatgg
     2341 aagttcctat actttctaga gaataggaac ttcatctttc tagaagatct cctacaatat
     2401 tctcagctgc catggaaaat cgatgaagga tcgcttgtct gggcaagagg atcaggatcg
     2461 ggatggtctt gattctgctg gaggaggagg aggagaagtc gaggaagcag cagcgaaagt
     2521 gcaagtgcga gtggtggaag tttggagtgc agcacaacaa aatcaacaac aacaccaact
     2581 acaagatgaa aagagcggaa ccacctgcac accatcatca ttatcatcat cgttttgggc
     2641 gcatgttgtg tggttccagc gtattaatat aattaattta ttccacatga gatatgatat
     2701 gatatactat gtattttttg tttttttttt atttgtaaac ctttaatata acaagaacta
     2761 caaaaaatga aaatgagcga aaatgcatat tctgccattc cacacacaca ccaacaacac
     2821 ccaacacacg cacacccaca agcttcagga aactgagaca ggaattttat taaaaattta
     2881 aattttgaag aaagttcagg gttaatagca tccatttttt gctttgcaag ttcctcagca
     2941 ttcttaacaa aagacgtctc ttttgacatg tttaaagttt aaacctcctg tgtgaaatta
     3001 ttatccgctc ataattccac acattatacg agccggaagc ataaagtgta aagcctgggg
     3061 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgccaa ttgctttcca
     3121 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3181 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3241 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3301 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3361 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3421 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3481 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3541 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3601 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3661 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3721 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3781 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3841 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3901 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3961 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4021 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4081 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4141 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4201 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4261 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4321 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4381 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4441 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4501 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4561 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4621 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4681 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4741 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4801 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4861 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4921 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4981 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5041 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5101 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5161 aacctataaa aataggcgta tcacgaggcc
//
