LOCUS       sequence-136540-        4073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for N-terminal Strep-8xHis-Strep-TEV site tag, inducible 
            Blasticidin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4073)
  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 4073)
  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: pKF290
FEATURES             Location/Qualifiers
     source          1..4073
                     /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"
     CDS             1342..1365
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1372..1395
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1408..1428
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        complement(join(1905..1911,1912..1929,1930..1935))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1950..1971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2091..2108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2262..2850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2342..2361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3021..3812,3813..3881))
                     /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     3644..3663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3882..3986)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4054..4072
                     /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 agaaaggtgg acatcaccac catcaccatc atcacggatc gtggagccac
     1381 ccgcagttcg aaaaaaccgg tgaattcgaa aacctgtatt ttcagggcga attcggtacc
     1441 tctccagctg ctccaagcgg cggtactagt atctttctag aagatctcct acaatattct
     1501 cagctgccat ggaaaatcga tgttcttctt ttattctctc aagattttca ggctgtatat
     1561 taaaacttat attaagaact atgctaacca cctcatcagg aaccgttgta ggtggcgtgg
     1621 gttttcttgg caatcgactc tcatgaaaac tacgagctaa atattcaata tgttcctctt
     1681 gaccaacttt attctgcatt ttttttgaac gaggtttaga gcaagcttca ggaaactgag
     1741 acaggaattt tattaaaaat ttaaattttg aagaaagttc agggttaata gcatccattt
     1801 tttgctttgc aagttcctca gcattcttaa caaaagacgt ctcttttgac atgtttaaag
     1861 tttaaacctc ctgtgtgaaa ttattatccg ctcataattc cacacattat acgagccgga
     1921 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1981 cgctcactgc caattgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2041 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2101 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2161 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2221 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2281 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2341 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2401 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2461 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2521 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2581 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2641 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2701 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2761 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2821 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2881 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2941 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3001 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3061 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3121 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3181 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3241 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3301 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3361 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3421 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3481 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3541 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3601 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3661 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3721 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3781 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3841 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3901 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3961 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4021 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gcc
//
