LOCUS       sequence-136534-        4053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for C-terminal TEV-site + V5-tag, Puromycin-Resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4053)
  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 4053)
  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: pSK32
FEATURES             Location/Qualifiers
     source          1..4053
                     /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             358..378
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             391..432
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(442..475)
                     /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(1417..1450)
                     /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(1885..1891,1892..1909,1910..1915))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1930..1951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2071..2088)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2242..2830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2322..2341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3001..3792,3793..3861))
                     /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     3624..3643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3862..3966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4034..4052
                     /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 gctcgaggaa
      361 aacctgtatt ttcagggcgg ttccggatct ggcaagccga tcccgaatcc gctgttaggc
      421 ctggatagca cctgaactag tgaagttcct atactttcta gagaatagga acttcaaata
      481 aacatatggg cgcgccctgt tggaatatac tattcaacct acaaaaataa cgttaaacaa
      541 cactacttta tatttgatat gaatggccac accttttatg ccataaaaca tattgtaaga
      601 gaataccact ctttttattc cttctttcct tcttgtacgt tttttgctgt gagtaggtcg
      661 tggtgctggt gttgcagttg aaataactta aaatataaat cataaaactc aaacataaac
      721 ttgactattt atttatttat taagaaagga ggtaccggtg gcgcgccatg accgagtaca
      781 agcctacagt gcgactggcc acacgcgatg acgttcccag agccgttcgg acgttggctg
      841 ccgcgtttgc ggactatccc gcaactcgcc atactgtgga ccctgatcgc cacatcgaga
      901 gggtgactga gcttcaggag ctgttcctga ccagggtagg gctggacata ggcaaggtct
      961 gggtggccga cgatggagcc gctgttgcgg tgtggactac cccagagagc gtggaagcag
     1021 gggctgtctt tgccgaaatt gggccaagga tggccgaact gtctgggtcc cgtttggctg
     1081 cccaacagca gatggaagga ctgcttgccc cacatcgacc caaggagcct gcatggttcc
     1141 tcgctaccgt gggtgtgagc ccagatcacc agggcaaagg tctgggatca gccgtcgttc
     1201 tgcctggcgt cgaggctgct gagagagcag gagtaccggc attcctcgaa accagtgctc
     1261 cacggaacct cccgttctac gagagactgg gctttacggt cacagccgat gtggaagtgc
     1321 ccgaaggccc tagaacctgg tgcatgacac ggaaacccgg tgcatgaaaa ttaaatataa
     1381 ataaagctat taaaataaat tactatctaa catatggaag ttcctatact ttctagagaa
     1441 taggaacttc atctttctag aagatctcct acaatattct cagctgccat ggaaaatcga
     1501 tgttcttctt ttattctctc aagattttca ggctgtatat taaaacttat attaagaact
     1561 atgctaacca cctcatcagg aaccgttgta ggtggcgtgg gttttcttgg caatcgactc
     1621 tcatgaaaac tacgagctaa atattcaata tgttcctctt gaccaacttt attctgcatt
     1681 ttttttgaac gaggtttaga gcaagcttca ggaaactgag acaggaattt tattaaaaat
     1741 ttaaattttg aagaaagttc agggttaata gcatccattt tttgctttgc aagttcctca
     1801 gcattcttaa caaaagacgt ctcttttgac atgtttaaag tttaaacctc ctgtgtgaaa
     1861 ttattatccg ctcataattc cacacattat acgagccgga agcataaagt gtaaagcctg
     1921 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc caattgcttt
     1981 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2041 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2101 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2641 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     2701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3061 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4021 attaacctat aaaaataggc gtatcacgag gcc
//
