LOCUS       sequence-136536-        3956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template base vector for N-terminal tagging, inducible Blasticidin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3956)
  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 3956)
  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: pKF292
FEATURES             Location/Qualifiers
     source          1..3956
                     /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"
     protein_bind    complement(372..404)
                     /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             complement(453..851)
                     /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(1254..1287)
                     /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(1788..1794,1795..1812,1813..1818))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1833..1854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1974..1991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2145..2733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2225..2244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2904..3695,3696..3764))
                     /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     3527..3546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3765..3869)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3937..3955
                     /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 gctcgagttt
      361 ttcagcaaga taagttccta tactttctag agaataggaa cttccatatg ttagaaacaa
      421 atttattttt aaagttttat ttttaataat ttctagccct cccacacata accagagggc
      481 agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg
      541 atcccaggat gcagatcgag aagaacctgt cggcaccgtc cgcaggggct caagatgccc
      601 ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca
      661 gtgcccagca ccacgagttc tgcgcaaggt cccccagtaa aatgatatac attgacacca
      721 gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg
      781 gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa
      841 ggcttggcca tatgcctagg cttgttgatt tgttttttgt ttatttactg cggtaattcc
      901 tagtcttaga agcttattga acttgttgat gccttctggt tgggaaatcc tcctctttat
      961 acccgatcca gtccaatcga aactgtgggc cgtggtgccc gataagataa gaaagccgtg
     1021 tgcaagtcga tcggcgtgcg caaaagcata atgctttttc tgcccggcga tgataagcga
     1081 tttgcacacg ccctgataac aggaattgat ttttttggtg cgcaaagcag tgaaagctct
     1141 ataagaatgc tggaccgcca gtcgacaatc atcaaacagt gctagctaac ggctaagtga
     1201 atcaagttta ttttaatata atcaacaaca aaatatttta acacaaacct agggaagttc
     1261 ctatactttc tagagaatag gaacttctaa tccaaaatgg gatccaccgg tgaattcggt
     1321 acctctccag ctgctccaag cggcggtact agtatctttc tagaagatct cctacaatat
     1381 tctcagctgc catggaaaat cgatgttctt cttttattct ctcaagattt tcaggctgta
     1441 tattaaaact tatattaaga actatgctaa ccacctcatc aggaaccgtt gtaggtggcg
     1501 tgggttttct tggcaatcga ctctcatgaa aactacgagc taaatattca atatgttcct
     1561 cttgaccaac tttattctgc attttttttg aacgaggttt agagcaagct tcaggaaact
     1621 gagacaggaa ttttattaaa aatttaaatt ttgaagaaag ttcagggtta atagcatcca
     1681 ttttttgctt tgcaagttcc tcagcattct taacaaaaga cgtctctttt gacatgttta
     1741 aagtttaaac ctcctgtgtg aaattattat ccgctcataa ttccacacat tatacgagcc
     1801 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1861 ttgcgctcac tgccaattgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1921 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2101 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2161 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2221 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2281 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2341 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2401 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2461 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2521 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2581 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2641 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2701 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2761 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2881 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2941 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3001 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3061 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3121 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3181 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3241 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3301 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3361 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3421 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3481 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3541 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3601 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3661 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3721 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3781 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3841 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3901 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcc
//
