LOCUS       sequence-141388-        3868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for C-terminal TEV-site + 2x V5-tag (short version), 
            Blasticidin-Resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3868)
  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 3868)
  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: pKF296
FEATURES             Location/Qualifiers
     source          1..3868
                     /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             358..378
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(475..508)
                     /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             785..1183
                     /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(1232..1265)
                     /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(1700..1706,1707..1724,1725..1730))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1745..1766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1886..1903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2057..2645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2137..2156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2816..3607,3608..3676))
                     /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     3439..3458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3677..3781)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3849..3867
                     /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 gccgccagat cttccggatg gctcgaggag
      361 aacctgtact tccagggcgg tgccagcatc ccaaaccccc tgttgggcct ggatagccca
      421 gcagctccta gcatccccaa cccactgctg ggtctggatg gcagctgaac tagtgaagtt
      481 cctatacttt ctagagaata ggaacttcaa ataaacatat gctgttggaa tatactattc
      541 aacctacaaa aataacgtta aacaacacta ctttatattt gatatgaatg gccacacctt
      601 ttatgccata aaacatattg taagagaata ccactctttt tattccttct ttccttcttg
      661 tacgtttttt gctgtgagta ggtcgtggtg ctggtgttgc agttgaaata acttaaaata
      721 taaatcataa aactcaaaca taaacttgac tatttattta tttattaaga aaggaggtac
      781 cggtatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
      841 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
      901 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gcgcagaact
      961 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
     1021 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg ttcttctcga
     1081 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
     1141 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc tagaaattat taaaaataaa
     1201 actttaaaaa taaatttgtt tctaacatat ggaagttcct atactttcta gagaatagga
     1261 acttcatctt tctagaagat ctcctacaat attctcagct gccatggaaa atcgatgttc
     1321 ttcttttatt ctctcaagat tttcaggctg tatattaaaa cttatattaa gaactatgct
     1381 aaccacctca tcaggaaccg ttgtaggtgg cgtgggtttt cttggcaatc gactctcatg
     1441 aaaactacga gctaaatatt caatatgttc ctcttgacca actttattct gcattttttt
     1501 tgaacgaggt ttagagcaag cttcaggaaa ctgagacagg aattttatta aaaatttaaa
     1561 ttttgaagaa agttcagggt taatagcatc cattttttgc tttgcaagtt cctcagcatt
     1621 cttaacaaaa gacgtctctt ttgacatgtt taaagtttaa acctcctgtg tgaaattatt
     1681 atccgctcat aattccacac attatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1741 cctaatgagt gagctaactc acattaattg cgttgcgctc actgccaatt gctttccagt
     1801 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1861 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1921 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1981 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2041 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2101 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2161 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2221 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2281 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2341 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2401 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2461 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     2521 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2581 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2641 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2701 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2761 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2821 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2881 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2941 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3001 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3061 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3121 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3181 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3241 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3301 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3361 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3421 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3481 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3541 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3601 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3661 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3721 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3781 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     3841 cctataaaaa taggcgtatc acgaggcc
//
