LOCUS       sequence-136538-        3986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for N-terminal 1x Strep tag, inducible Blasticidin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3986)
  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 3986)
  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: pKF294
FEATURES             Location/Qualifiers
     source          1..3986
                     /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)"
     CDS             1312..1335
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(join(1818..1824,1825..1842,1843..1848))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1863..1884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2004..2021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2175..2763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2255..2274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2934..3725,3726..3794))
                     /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     3557..3576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3795..3899)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3967..3985
                     /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 gatccagcgc ttggagccac
     1321 ccgcagttcg aaaaaaccgg tgaattcggt acctctccag ctgctccaag cggcggtact
     1381 agtatctttc tagaagatct cctacaatat tctcagctgc catggaaaat cgatgttctt
     1441 cttttattct ctcaagattt tcaggctgta tattaaaact tatattaaga actatgctaa
     1501 ccacctcatc aggaaccgtt gtaggtggcg tgggttttct tggcaatcga ctctcatgaa
     1561 aactacgagc taaatattca atatgttcct cttgaccaac tttattctgc attttttttg
     1621 aacgaggttt agagcaagct tcaggaaact gagacaggaa ttttattaaa aatttaaatt
     1681 ttgaagaaag ttcagggtta atagcatcca ttttttgctt tgcaagttcc tcagcattct
     1741 taacaaaaga cgtctctttt gacatgttta aagtttaaac ctcctgtgtg aaattattat
     1801 ccgctcataa ttccacacat tatacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1861 taatgagtga gctaactcac attaattgcg ttgcgctcac tgccaattgc tttccagtcg
     1921 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1981 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2041 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2101 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2161 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2221 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2281 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2341 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2401 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2461 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2521 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2581 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     2641 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2701 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2761 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2821 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2881 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2941 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3001 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3061 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3121 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3181 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3241 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3301 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3361 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3421 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3481 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3541 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3601 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3661 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3721 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3781 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3841 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3901 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3961 tataaaaata ggcgtatcac gaggcc
//
