LOCUS       sequence-229440-        2183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes sgRNA targeting position 9 in pColE1_70a_deGFP_KanR.
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_pos9_AmpR
FEATURES             Location/Qualifiers
     source          1..2183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..76
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        97..172
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(join(434..1225,1226..1294))
                     /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     1057..1076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1295..1399)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1502..2090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1991..2010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaccgtgcgg cccgagagca tcgatcctag catggatgca tttgacagct agctcagtcc
       61 taggtataat gctagcgtcg ccctcgaact tcacctgttt tagagctaga aatagcaagt
      121 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttc
      181 cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt
      241 ttggaggggc agaaagatga atgactgtcc acgacgctat acccaaaaga aaaagcttga
      301 aagaatcgat cgatcccgaa tgagtagtac agcggccgcg attatcaaaa aggatcttca
      361 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
      421 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      481 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      541 taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg gctccagatt
      601 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
      661 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
      721 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
      781 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
      841 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
      901 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
      961 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1021 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1081 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1141 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1201 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1261 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1321 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1381 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgtcatg accaaaatcc
     1441 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1501 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1561 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1621 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     1681 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1741 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1801 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1861 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     1921 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     1981 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2041 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2101 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2161 cgttatcccc tgattctgtg gat
//
