LOCUS       sequence-136532-        4068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for C-terminal Twin-Strep tag, Puromycin-resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4068)
  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 4068)
  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: pSK24
FEATURES             Location/Qualifiers
     source          1..4068
                     /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             364..387
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             424..447
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    complement(457..490)
                     /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)"
     protein_bind    complement(1432..1465)
                     /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(1900..1906,1907..1924,1925..1930))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1945..1966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2086..2103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2257..2845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2337..2356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3016..3807,3808..3876))
                     /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     3639..3658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3877..3981)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4049..4067
                     /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 gctcgagagc
      361 gcttggagcc acccgcagtt cgagaaaggt ggaggttccg gaggtggatc gggaggtgga
      421 tcgtggagcc acccgcagtt cgaaaaatga actagtgaag ttcctatact ttctagagaa
      481 taggaacttc aaataaacat atgggcgcgc cctgttggaa tatactattc aacctacaaa
      541 aataacgtta aacaacacta ctttatattt gatatgaatg gccacacctt ttatgccata
      601 aaacatattg taagagaata ccactctttt tattccttct ttccttcttg tacgtttttt
      661 gctgtgagta ggtcgtggtg ctggtgttgc agttgaaata acttaaaata taaatcataa
      721 aactcaaaca taaacttgac tatttattta tttattaaga aaggaggtac cggtggcgcg
      781 ccatgaccga gtacaagcct acagtgcgac tggccacacg cgatgacgtt cccagagccg
      841 ttcggacgtt ggctgccgcg tttgcggact atcccgcaac tcgccatact gtggaccctg
      901 atcgccacat cgagagggtg actgagcttc aggagctgtt cctgaccagg gtagggctgg
      961 acataggcaa ggtctgggtg gccgacgatg gagccgctgt tgcggtgtgg actaccccag
     1021 agagcgtgga agcaggggct gtctttgccg aaattgggcc aaggatggcc gaactgtctg
     1081 ggtcccgttt ggctgcccaa cagcagatgg aaggactgct tgccccacat cgacccaagg
     1141 agcctgcatg gttcctcgct accgtgggtg tgagcccaga tcaccagggc aaaggtctgg
     1201 gatcagccgt cgttctgcct ggcgtcgagg ctgctgagag agcaggagta ccggcattcc
     1261 tcgaaaccag tgctccacgg aacctcccgt tctacgagag actgggcttt acggtcacag
     1321 ccgatgtgga agtgcccgaa ggccctagaa cctggtgcat gacacggaaa cccggtgcat
     1381 gaaaattaaa tataaataaa gctattaaaa taaattacta tctaacatat ggaagttcct
     1441 atactttcta gagaatagga acttcatctt tctagaagat ctcctacaat attctcagct
     1501 gccatggaaa atcgatgttc ttcttttatt ctctcaagat tttcaggctg tatattaaaa
     1561 cttatattaa gaactatgct aaccacctca tcaggaaccg ttgtaggtgg cgtgggtttt
     1621 cttggcaatc gactctcatg aaaactacga gctaaatatt caatatgttc ctcttgacca
     1681 actttattct gcattttttt tgaacgaggt ttagagcaag cttcaggaaa ctgagacagg
     1741 aattttatta aaaatttaaa ttttgaagaa agttcagggt taatagcatc cattttttgc
     1801 tttgcaagtt cctcagcatt cttaacaaaa gacgtctctt ttgacatgtt taaagtttaa
     1861 acctcctgtg tgaaattatt atccgctcat aattccacac attatacgag ccggaagcat
     1921 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1981 actgccaatt gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2041 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2101 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2161 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2221 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2281 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2341 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2401 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2461 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2521 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2581 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2641 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2701 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2761 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2821 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2881 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2941 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3001 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3061 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3121 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3181 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3241 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3301 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3361 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3421 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3481 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3541 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3601 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3661 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3721 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3781 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3841 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3901 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3961 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4021 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcc
//
