LOCUS       sequence-136544-        4285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for N-terminal 3x FLAG tag, constitutive Blasticidin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4285)
  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 4285)
  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: pRB34
FEATURES             Location/Qualifiers
     source          1..4285
                     /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(372..404)
                     /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             674..1072
                     /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(1520..1553)
                     /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(2117..2123,2124..2141,2142..2147))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2162..2183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2303..2320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2474..3062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2554..2573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3233..4024,4025..4093))
                     /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     3856..3875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4094..4198)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4266..4284
                     /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 gctcgagttt
      361 ttcagcaaga taagttccta tactttctag agaataggaa cttccatatg ctgttggaat
      421 atactattca acctacaaaa ataacgttaa acaacactac tttatatttg atatgaatgg
      481 ccacaccttt tatgccataa aacatattgt aagagaatac cactcttttt attccttctt
      541 tccttcttgt acgttttttg ctgtgagtag gtcgtggtgc tggtgttgca gttgaaataa
      601 cttaaaatat aaatcataaa actcaaacat aaacttgact atttatttat ttattaagaa
      661 aggaggtacc ggtatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
      721 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
      781 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
      841 cgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
      901 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
      961 tcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     1021 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct agaaattatt
     1081 aaaaataaaa ctttaaaaat aaatttgttt ctaacatatg cctaggcttg ttgatttgtt
     1141 ttttgtttat ttactgcggt aattcctagt cttagaagct tattgaactt gttgatgcct
     1201 tctggttggg aaatcctcct ctttataccc gatccagtcc aatcgaaact gtgggccgtg
     1261 gtgcccgata agataagaaa gccgtgtgca agtcgatcgg cgtgcgcaaa agcataatgc
     1321 tttttctgcc cggcgatgat aagcgatttg cacacgccct gataacagga attgattttt
     1381 ttggtgcgca aagcagtgaa agctctataa gaatgctgga ccgccagtcg acaatcatca
     1441 aacagtgcta gctaacggct aagtgaatca agtttatttt aatataatca acaacaaaat
     1501 attttaacac aaacctaggg aagttcctat actttctaga gaataggaac ttctaatcca
     1561 aaatgggatc caccggtgaa ttcgactaca aggaccacga cggcgactac aaggaccacg
     1621 acatcgacta caaggacgac gacaagggta cctctccagc tgctccaagc ggcggtacta
     1681 gtatctttct agaagatctc ctacaatatt ctcagctgcc atggaaaatc gatgttcttc
     1741 ttttattctc tcaagatttt caggctgtat attaaaactt atattaagaa ctatgctaac
     1801 cacctcatca ggaaccgttg taggtggcgt gggttttctt ggcaatcgac tctcatgaaa
     1861 actacgagct aaatattcaa tatgttcctc ttgaccaact ttattctgca ttttttttga
     1921 acgaggttta gagcaagctt caggaaactg agacaggaat tttattaaaa atttaaattt
     1981 tgaagaaagt tcagggttaa tagcatccat tttttgcttt gcaagttcct cagcattctt
     2041 aacaaaagac gtctcttttg acatgtttaa agtttaaacc tcctgtgtga aattattatc
     2101 cgctcataat tccacacatt atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2161 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gccaattgct ttccagtcgg
     2221 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2281 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2341 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2401 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2461 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2521 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2581 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2641 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2701 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2761 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2821 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2881 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2941 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3001 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3061 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3121 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3181 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3241 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3301 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3361 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3421 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3481 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3541 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3601 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3661 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3721 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3781 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3841 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3901 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3961 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4021 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4081 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4141 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4201 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4261 ataaaaatag gcgtatcacg aggcc
//
