LOCUS       sequence-136545-        4252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for N-terminal 1x Strep tag, constitutive Blasticidin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4252)
  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 4252)
  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: pRB32
FEATURES             Location/Qualifiers
     source          1..4252
                     /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)"
     CDS             1578..1601
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(join(2084..2090,2091..2108,2109..2114))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2129..2150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2270..2287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2441..3029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2521..2540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3200..3991,3992..4060))
                     /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     3823..3842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4061..4165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4233..4251
                     /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 cagcgcttgg agccacccgc agttcgaaaa aaccggtgaa ttcggtacct
     1621 ctccagctgc tccaagcggc ggtactagta tctttctaga agatctccta caatattctc
     1681 agctgccatg gaaaatcgat gttcttcttt tattctctca agattttcag gctgtatatt
     1741 aaaacttata ttaagaacta tgctaaccac ctcatcagga accgttgtag gtggcgtggg
     1801 ttttcttggc aatcgactct catgaaaact acgagctaaa tattcaatat gttcctcttg
     1861 accaacttta ttctgcattt tttttgaacg aggtttagag caagcttcag gaaactgaga
     1921 caggaatttt attaaaaatt taaattttga agaaagttca gggttaatag catccatttt
     1981 ttgctttgca agttcctcag cattcttaac aaaagacgtc tcttttgaca tgtttaaagt
     2041 ttaaacctcc tgtgtgaaat tattatccgc tcataattcc acacattata cgagccggaa
     2101 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2161 gctcactgcc aattgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2221 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2281 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2341 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2401 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2461 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2521 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2581 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2641 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2701 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2761 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2821 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2881 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2941 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3001 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3061 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3121 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3181 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3241 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3301 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3361 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3421 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3481 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3541 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3601 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3661 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3721 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3781 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3841 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3901 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3961 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4021 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4081 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4141 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     4201 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg cc
//
