LOCUS       sequence-136307-        4745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  serves as template for PCR-mediated fusion of U6-promoter with 
            sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4745)
  AUTHORS   Bottcher R, Hollmann M, Merk K, Nitschko V, Obermaier C, 
            Philippou-Massier J, Wieland I, Gaul U, Forstemann K
  TITLE     Efficient chromosomal gene modification with CRISPR/cas9 and 
            PCR-based homologous recombination donors in cultured Drosophila 
            cells.
  JOURNAL   Nucleic Acids Res. 2014 Apr 19.
  PUBMED    24748663
REFERENCE   2  (bases 1 to 4745)
  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"; journalName: "Nucleic 
            Acids Res. 2014 Apr 19."
COMMENT     Sequence Label: pRB17
FEATURES             Location/Qualifiers
     source          1..4745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(1239..1245,1246..1263,1264..1269))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2622..2643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2763..2780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2934..3522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3014..3033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3693..4484,4485..4553))
                     /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     4316..4335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4554..4658)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4726..4744
                     /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 tgctcacctg tgattgctcc tactcaaata caaaaacatc aaattttctg
      421 tcaataaagc atatttattt atatttattt tacaggaaag aattcctttt aaagtgtatt
      481 ttaacctata atgaaaaacg attaaaaaaa atacataaaa taattcgaaa atttttgaat
      541 agcccaggtt gataaaaatt catttcatac gttttataac ttatgcccct aagtattttt
      601 tgaccatagt gtttcaattc tacattaatt ttacagagta gaatgaaacg ccacctactc
      661 agccaagagg cgaaaaggtt agctcgccaa gcagagaggg cgccagtgct cactactttt
      721 tataattctc aacttctttt tccagactca gttcgtatat atagacctat tttcaattta
      781 acgtcgttta atacgactca ctatatcttt ctagaagatc tcctacaata ttctcagctg
      841 ccatggaaaa tcgatgttct tcttttattc tctcaagatt ttcaggctgt atattaaaac
      901 ttatattaag aactatgcta accacctcat caggaaccgt tgtaggtggc gtgggttttc
      961 ttggcaatcg actctcatga aaactacgag ctaaatattc aatatgttcc tcttgaccaa
     1021 ctttattctg catttttttt gaacgaggtt tagagcaagc ttcaggaaac tgagacagga
     1081 attttattaa aaatttaaat tttgaagaaa gttcagggtt aatagcatcc attttttgct
     1141 ttgcaagttc ctcagcattc ttaacaaaag acgtctcttt tgacatgttt aaagtttaaa
     1201 cctcctgtgt gaaattatta tccgctcata attccacaca ttatacgagc cggaagcata
     1261 aagtgtaaag cctggggtgc ctaatgctga tgaatcccct aatgattttt atcaaaatca
     1321 ttaagttaag gtagatacac atcttgtcat atgatcaaat ggtttcgcca aaaatcaata
     1381 atcagacaac aaaatgtgcg aactcgatat tttacacgac tctctttacc aattctgccc
     1441 cgaattacac ttaaaacgac tcaacagctt aacgttggct tgccacgcct tacttgactg
     1501 taaaactctc actcttaccg aacttggccg taacctgcca accaaagcga gaacaaaaca
     1561 taacatcaaa cgaatcgacc gattgttagg taatcgtcac ctccacaaag agcgactcgc
     1621 tgtataccgt tggcatgcta gctttatctg ttcgggcaat acgatgccca ttgtacttgt
     1681 tgactggtct gatatccgtg agcaaaaacg gcttatggta ttgcgagctt cagtcgcact
     1741 acacggtcgt tctgttactc tttatgagaa agcgttcccg ctttcagagc aatgttcaaa
     1801 gaaagctcat gaccaatttc tagccgacct tgcgagcatt ctaccgagta acaccacacc
     1861 gctcattgtc agtgatgctg gctttaaagt gccatggtat aaatccgttg aaaagctggg
     1921 ttggtactgg ttaagtcgag taaaaggaaa agtacaatat gcagacctag gagcggaaaa
     1981 ctggaaacct atcagcaact tacatgatat gtcatctagt cactcaaaga ctttaggcta
     2041 taagaggctg actaaaagca atccaatctc atgccaaatt ctattgtata aatctcgctc
     2101 taaaggccga aaaaatcagc gctcgacacg gactcattgt caccacccgt cacctaaaat
     2161 ctactcagcg tcggcaaagg agccatggat tctagcaact aacttacctg ttgaaattcg
     2221 aacacccaaa caacttgtta atatctattc gaagcgaatg cagattgaag aaaccttccg
     2281 agacttgaaa agtcctgcct acggactagg cctacgccat agccgaacga gcagctcaga
     2341 gcgttttgat atcatgctgc taatcgccct gatgcttcaa ctaacatgtt ggcttgcggg
     2401 cgttcatgct cagaaacaag gttgggacaa gcacttccag gctaacacag tcagaaatcg
     2461 aaacgtactc tcaacagttc gcttaggcat ggaagttttg cggcattctg gctacacaat
     2521 aacaagggaa gacttactcg tggctgcaac cctactagct caaaatttat tcacacatgg
     2581 ttacgctttg gggaaattat gaggggatct ctcagtgcct aatgagtgag ctaactcaca
     2641 ttaattgcgt tgcgctcact gccaattgct ttccagtcgg gaaacctgtc gtgccagctg
     2701 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2761 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2821 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3361 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4021 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4081 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4141 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4201 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4261 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4321 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4381 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4441 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4501 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4561 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4621 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4681 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4741 aggcc
//
