LOCUS       sequence-141561-        4014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for insertion of attP site (C-terminal tag plasmid design),
            Puromycin-Resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4014)
  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 4014)
  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: pSK41
FEATURES             Location/Qualifiers
     source          1..4014
                     /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)"
     protein_bind    complement(1378..1411)
                     /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(1846..1852,1853..1870,1871..1876))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1891..1912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2032..2049)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2203..2791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2283..2302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2962..3753,3754..3822))
                     /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     3585..3604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3823..3927)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3995..4013
                     /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 aaacatatgg gcgcgccctg ttggaatata ctattcaacc
      481 tacaaaaata acgttaaaca acactacttt atatttgata tgaatggcca caccttttat
      541 gccataaaac atattgtaag agaataccac tctttttatt ccttctttcc ttcttgtacg
      601 ttttttgctg tgagtaggtc gtggtgctgg tgttgcagtt gaaataactt aaaatataaa
      661 tcataaaact caaacataaa cttgactatt tatttattta ttaagaaagg aggtaccggt
      721 ggcgcgccat gaccgagtac aagcctacag tgcgactggc cacacgcgat gacgttccca
      781 gagccgttcg gacgttggct gccgcgtttg cggactatcc cgcaactcgc catactgtgg
      841 accctgatcg ccacatcgag agggtgactg agcttcagga gctgttcctg accagggtag
      901 ggctggacat aggcaaggtc tgggtggccg acgatggagc cgctgttgcg gtgtggacta
      961 ccccagagag cgtggaagca ggggctgtct ttgccgaaat tgggccaagg atggccgaac
     1021 tgtctgggtc ccgtttggct gcccaacagc agatggaagg actgcttgcc ccacatcgac
     1081 ccaaggagcc tgcatggttc ctcgctaccg tgggtgtgag cccagatcac cagggcaaag
     1141 gtctgggatc agccgtcgtt ctgcctggcg tcgaggctgc tgagagagca ggagtaccgg
     1201 cattcctcga aaccagtgct ccacggaacc tcccgttcta cgagagactg ggctttacgg
     1261 tcacagccga tgtggaagtg cccgaaggcc ctagaacctg gtgcatgaca cggaaacccg
     1321 gtgcatgaaa attaaatata aataaagcta ttaaaataaa ttactatcta acatatggaa
     1381 gttcctatac tttctagaga ataggaactt catctttcta gaagatctcc tacaatattc
     1441 tcagctgcca tggaaaatcg atgttcttct tttattctct caagattttc aggctgtata
     1501 ttaaaactta tattaagaac tatgctaacc acctcatcag gaaccgttgt aggtggcgtg
     1561 ggttttcttg gcaatcgact ctcatgaaaa ctacgagcta aatattcaat atgttcctct
     1621 tgaccaactt tattctgcat tttttttgaa cgaggtttag agcaagcttc aggaaactga
     1681 gacaggaatt ttattaaaaa tttaaatttt gaagaaagtt cagggttaat agcatccatt
     1741 ttttgctttg caagttcctc agcattctta acaaaagacg tctcttttga catgtttaaa
     1801 gtttaaacct cctgtgtgaa attattatcc gctcataatt ccacacatta tacgagccgg
     1861 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1921 gcgctcactg ccaattgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1981 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2041 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2101 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2161 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2221 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2281 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2341 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2401 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2461 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2521 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2581 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2641 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2701 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2761 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2821 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2881 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2941 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3001 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3061 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3121 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3181 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3241 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3301 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3361 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3421 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3481 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3541 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3601 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3661 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3721 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3781 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3841 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3901 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3961 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggcc
//
