LOCUS       sequence-136535-        3796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for insertion of attP site (C-terminal tag plasmid design),
            Blasticidin-Resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3796)
  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 3796)
  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: pSK15
FEATURES             Location/Qualifiers
     source          1..3796
                     /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(403..436)
                     /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             713..1111
                     /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(1160..1193)
                     /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(1628..1634,1635..1652,1653..1658))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1673..1694
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1814..1831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1985..2573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2065..2084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2744..3535,3536..3604))
                     /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     3367..3386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3605..3709)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3777..3795
                     /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 gctcgagccc
      361 caactggggt aacctttgag ttctctcagt tgggggacta gtgaagttcc tatactttct
      421 agagaatagg aacttcaaat aaacatatgc tgttggaata tactattcaa cctacaaaaa
      481 taacgttaaa caacactact ttatatttga tatgaatggc cacacctttt atgccataaa
      541 acatattgta agagaatacc actcttttta ttccttcttt ccttcttgta cgttttttgc
      601 tgtgagtagg tcgtggtgct ggtgttgcag ttgaaataac ttaaaatata aatcataaaa
      661 ctcaaacata aacttgacta tttatttatt tattaagaaa ggaggtaccg gtatggccaa
      721 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
      781 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
      841 cactggtgtc aatgtatatc attttactgg gggaccttgc gcagaactcg tggtgctggg
      901 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
      961 caggggcatc ttgagcccct gcggacggtg ccgacaggtt cttctcgatc tgcatcctgg
     1021 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     1081 gctgccctct ggttatgtgt gggagggcta gaaattatta aaaataaaac tttaaaaata
     1141 aatttgtttc taacatatgg aagttcctat actttctaga gaataggaac ttcatctttc
     1201 tagaagatct cctacaatat tctcagctgc catggaaaat cgatgttctt cttttattct
     1261 ctcaagattt tcaggctgta tattaaaact tatattaaga actatgctaa ccacctcatc
     1321 aggaaccgtt gtaggtggcg tgggttttct tggcaatcga ctctcatgaa aactacgagc
     1381 taaatattca atatgttcct cttgaccaac tttattctgc attttttttg aacgaggttt
     1441 agagcaagct tcaggaaact gagacaggaa ttttattaaa aatttaaatt ttgaagaaag
     1501 ttcagggtta atagcatcca ttttttgctt tgcaagttcc tcagcattct taacaaaaga
     1561 cgtctctttt gacatgttta aagtttaaac ctcctgtgtg aaattattat ccgctcataa
     1621 ttccacacat tatacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1681 gctaactcac attaattgcg ttgcgctcac tgccaattgc tttccagtcg ggaaacctgt
     1741 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1801 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1861 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1921 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2161 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2401 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     2461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2881 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3121 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3721 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3781 ggcgtatcac gaggcc
//
