LOCUS       sequence-229416-        2183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes non-targeting sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2183)
  AUTHORS   Collias D, Marshall R, Collins SP, Beisel CL, Noireaux V
  TITLE     An educational module to explore CRISPR technologies with a 
            cell-free transcription-translation system.
  JOURNAL   Synth Biol (Oxf). 2019 Jan 21;4(1):ysz005. doi: 
            10.1093/synbio/ysz005. eCollection 2019.
  PUBMED    32995532
REFERENCE   2  (bases 1 to 2183)
  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: "Synth Biol 
            (Oxf)."; date: "2019-01-21"; volume: "4(1):ysz005. doi"; pages: " 
            10.1093/synbio/ysz005. eCollection 2019"
COMMENT     Sequence Label: pColE1_sgRNA_NT_AmpR
FEATURES             Location/Qualifiers
     source          1..2183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        51..126
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(join(388..1179,1180..1248))
                     /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     1011..1030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1249..1353)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1456..2044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1945..1964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2179..2183,1..30)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 agctagctca gtcctaggta taatgctagc aagcagatta cgttcaagca gttttagagc
       61 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      121 cggtgctttt tttccggctt atcggtcagt ttcacctgat ttacgtaaaa acccgcttcg
      181 gcgggttttt gcttttggag gggcagaaag atgaatgact gtccacgacg ctatacccaa
      241 aagaaaaagc ttgaaagaat cgatcgatcc cgaatgagta gtacagcggc cgcgattatc
      301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
      541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
      601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
      661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
      721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
      841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
      901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
      961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgt
     1381 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1441 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1501 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1561 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1621 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1681 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1741 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1801 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1861 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1921 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1981 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2041 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2101 catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccc gagagcatcg
     2161 atcctagcat ggatgcattt gac
//
