LOCUS       sequence-141389-        4086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for C-terminal TEV-site + 2x V5-tag (short version), 
            Puromycin-Resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4086)
  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 4086)
  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: pKF297
FEATURES             Location/Qualifiers
     source          1..4086
                     /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"
     protein_bind    complement(475..508)
                     /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(1450..1483)
                     /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(1918..1924,1925..1942,1943..1948))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1963..1984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2104..2121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2275..2863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2355..2374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3034..3825,3826..3894))
                     /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     3657..3676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3895..3999)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4067..4085
                     /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 gctcgaggag
      361 aacctgtact tccagggcgg tgccagcatc ccaaaccccc tgttgggcct ggatagccca
      421 gcagctccta gcatccccaa cccactgctg ggtctggatg gcagctgaac tagtgaagtt
      481 cctatacttt ctagagaata ggaacttcaa ataaacatat gggcgcgccc tgttggaata
      541 tactattcaa cctacaaaaa taacgttaaa caacactact ttatatttga tatgaatggc
      601 cacacctttt atgccataaa acatattgta agagaatacc actcttttta ttccttcttt
      661 ccttcttgta cgttttttgc tgtgagtagg tcgtggtgct ggtgttgcag ttgaaataac
      721 ttaaaatata aatcataaaa ctcaaacata aacttgacta tttatttatt tattaagaaa
      781 ggaggtaccg gtggcgcgcc atgaccgagt acaagcctac agtgcgactg gccacacgcg
      841 atgacgttcc cagagccgtt cggacgttgg ctgccgcgtt tgcggactat cccgcaactc
      901 gccatactgt ggaccctgat cgccacatcg agagggtgac tgagcttcag gagctgttcc
      961 tgaccagggt agggctggac ataggcaagg tctgggtggc cgacgatgga gccgctgttg
     1021 cggtgtggac taccccagag agcgtggaag caggggctgt ctttgccgaa attgggccaa
     1081 ggatggccga actgtctggg tcccgtttgg ctgcccaaca gcagatggaa ggactgcttg
     1141 ccccacatcg acccaaggag cctgcatggt tcctcgctac cgtgggtgtg agcccagatc
     1201 accagggcaa aggtctggga tcagccgtcg ttctgcctgg cgtcgaggct gctgagagag
     1261 caggagtacc ggcattcctc gaaaccagtg ctccacggaa cctcccgttc tacgagagac
     1321 tgggctttac ggtcacagcc gatgtggaag tgcccgaagg ccctagaacc tggtgcatga
     1381 cacggaaacc cggtgcatga aaattaaata taaataaagc tattaaaata aattactatc
     1441 taacatatgg aagttcctat actttctaga gaataggaac ttcatctttc tagaagatct
     1501 cctacaatat tctcagctgc catggaaaat cgatgttctt cttttattct ctcaagattt
     1561 tcaggctgta tattaaaact tatattaaga actatgctaa ccacctcatc aggaaccgtt
     1621 gtaggtggcg tgggttttct tggcaatcga ctctcatgaa aactacgagc taaatattca
     1681 atatgttcct cttgaccaac tttattctgc attttttttg aacgaggttt agagcaagct
     1741 tcaggaaact gagacaggaa ttttattaaa aatttaaatt ttgaagaaag ttcagggtta
     1801 atagcatcca ttttttgctt tgcaagttcc tcagcattct taacaaaaga cgtctctttt
     1861 gacatgttta aagtttaaac ctcctgtgtg aaattattat ccgctcataa ttccacacat
     1921 tatacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1981 attaattgcg ttgcgctcac tgccaattgc tttccagtcg ggaaacctgt cgtgccagct
     2041 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2101 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2161 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2461 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2701 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2881 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2941 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3001 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3061 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3121 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3181 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3241 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3301 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3361 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3421 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3481 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3541 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3601 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3661 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3721 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3781 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3841 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3901 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3961 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4021 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4081 gaggcc
//
