LOCUS       sequence-266843-        2469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Subcloning and expression of non-targeting sgRNA for use in yeast 
            dual reporter system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2469)
  AUTHORS   Standage-Beier K, Brookhouser N, Balachandran P, Zhang Q, Brafman 
            DA, Wang X
  TITLE     RNA-Guided Recombinase-Cas9 Fusion Targets Genomic DNA Deletion and 
            Integration.
  JOURNAL   CRISPR J. 2019 Aug;2:209-222. doi: 10.1089/crispr.2019.0013.
  PUBMED    31436506
REFERENCE   2  (bases 1 to 2469)
  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: "CRISPR J.";
            date: "2019-08"; volume: "2:209-222. doi"; pages: " 
            10.1089/crispr.2019.0013"
COMMENT     Sequence Label: pSB1C3-ysg(NT)
FEATURES             Location/Qualifiers
     source          1..2469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..275
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        294..369
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      443..500
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(524..541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(695..1283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(775..794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1466..1560)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1581..2240)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2155..2174
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2402..2420
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2424..2467)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattctctt tgaaaagata atgtatgatt atgctttcac tcatatttat acagaaactt
       61 gatgttttct ttcgagtata tacaaggtga ttacatgtac gtttgaagta caactctaga
      121 ttttgtagtg ccctcttggg ctagcggtaa aggtgcgcat tttttcacac cctacaatgt
      181 tctgttcaaa agattttggt caaacgctgt agaagtgaaa gttggtgcgc atgtttcggc
      241 gttcgaaact tctccgcagt gaaagataaa tgatcagaag agcgagctct tctgttttag
      301 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      361 agtcggtgct ttttttacta gataattgga gaccgagctg gtctctctag gatgtgcact
      421 agtagagctg cggccgctgc agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt
      481 tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc
      541 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      601 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      661 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca caggctccgc ccccctgacg
      721 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      781 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      841 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      901 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      961 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1021 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1081 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1141 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1201 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1261 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1321 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1381 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1441 cttggtctga cagctcgagg cttggattct caccaataaa aaacgcccgg cggcaaccga
     1501 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     1561 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     1621 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     1681 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     1741 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     1801 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     1861 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     1921 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     1981 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
     2041 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     2101 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     2161 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     2221 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     2281 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgcccg
     2341 atcaactcga gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     2401 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     2461 tgatttctg
//
