LOCUS       sequence-136550-        4470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template for N-terminal 1x Strep tag, constitutive Puromycin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4470)
  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 4470)
  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: pRB31
FEATURES             Location/Qualifiers
     source          1..4470
                     /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)"
     protein_bind    complement(1738..1771)
                     /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             1796..1819
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(join(2302..2308,2309..2326,2327..2332))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2347..2368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2488..2505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2659..3247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2739..2758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3418..4209,4210..4278))
                     /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     4041..4060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4279..4383)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4451..4469
                     /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 ggcgcgccct
      421 gttggaatat actattcaac ctacaaaaat aacgttaaac aacactactt tatatttgat
      481 atgaatggcc acacctttta tgccataaaa catattgtaa gagaatacca ctctttttat
      541 tccttctttc cttcttgtac gttttttgct gtgagtaggt cgtggtgctg gtgttgcagt
      601 tgaaataact taaaatataa atcataaaac tcaaacataa acttgactat ttatttattt
      661 attaagaaag gaggtaccgg tggcgcgcca tgaccgagta caagcctaca gtgcgactgg
      721 ccacacgcga tgacgttccc agagccgttc ggacgttggc tgccgcgttt gcggactatc
      781 ccgcaactcg ccatactgtg gaccctgatc gccacatcga gagggtgact gagcttcagg
      841 agctgttcct gaccagggta gggctggaca taggcaaggt ctgggtggcc gacgatggag
      901 ccgctgttgc ggtgtggact accccagaga gcgtggaagc aggggctgtc tttgccgaaa
      961 ttgggccaag gatggccgaa ctgtctgggt cccgtttggc tgcccaacag cagatggaag
     1021 gactgcttgc cccacatcga cccaaggagc ctgcatggtt cctcgctacc gtgggtgtga
     1081 gcccagatca ccagggcaaa ggtctgggat cagccgtcgt tctgcctggc gtcgaggctg
     1141 ctgagagagc aggagtaccg gcattcctcg aaaccagtgc tccacggaac ctcccgttct
     1201 acgagagact gggctttacg gtcacagccg atgtggaagt gcccgaaggc cctagaacct
     1261 ggtgcatgac acggaaaccc ggtgcatgaa aattaaatat aaataaagct attaaaataa
     1321 attactatct aacatatgcc taggcttgtt gatttgtttt ttgtttattt actgcggtaa
     1381 ttcctagtct tagaagctta ttgaacttgt tgatgccttc tggttgggaa atcctcctct
     1441 ttatacccga tccagtccaa tcgaaactgt gggccgtggt gcccgataag ataagaaagc
     1501 cgtgtgcaag tcgatcggcg tgcgcaaaag cataatgctt tttctgcccg gcgatgataa
     1561 gcgatttgca cacgccctga taacaggaat tgattttttt ggtgcgcaaa gcagtgaaag
     1621 ctctataaga atgctggacc gccagtcgac aatcatcaaa cagtgctagc taacggctaa
     1681 gtgaatcaag tttattttaa tataatcaac aacaaaatat tttaacacaa acctagggaa
     1741 gttcctatac tttctagaga ataggaactt ctaatccaaa atgggatcca gcgcttggag
     1801 ccacccgcag ttcgaaaaaa ccggtgaatt cggtacctct ccagctgctc caagcggcgg
     1861 tactagtatc tttctagaag atctcctaca atattctcag ctgccatgga aaatcgatgt
     1921 tcttctttta ttctctcaag attttcaggc tgtatattaa aacttatatt aagaactatg
     1981 ctaaccacct catcaggaac cgttgtaggt ggcgtgggtt ttcttggcaa tcgactctca
     2041 tgaaaactac gagctaaata ttcaatatgt tcctcttgac caactttatt ctgcattttt
     2101 tttgaacgag gtttagagca agcttcagga aactgagaca ggaattttat taaaaattta
     2161 aattttgaag aaagttcagg gttaatagca tccatttttt gctttgcaag ttcctcagca
     2221 ttcttaacaa aagacgtctc ttttgacatg tttaaagttt aaacctcctg tgtgaaatta
     2281 ttatccgctc ataattccac acattatacg agccggaagc ataaagtgta aagcctgggg
     2341 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgccaa ttgctttcca
     2401 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2461 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2521 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2581 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2641 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2701 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2761 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2821 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2881 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2941 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3001 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3061 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3121 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3181 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3241 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3301 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3361 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3421 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3481 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3541 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3601 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3661 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3721 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3781 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3841 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3901 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3961 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4021 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4081 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4141 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4201 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4261 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4381 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4441 aacctataaa aataggcgta tcacgaggcc
//
