LOCUS       sequence-225356-        2176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes sgRNA targeting position 6 in pColE1_70a_deGFP_KanR, 
            truncated in the 3' end.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2176)
  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 2176)
  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_T1_AmpR
FEATURES             Location/Qualifiers
     source          1..2176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     250..269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        484..518
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        539..614
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(join(869..1660,1661..1729))
                     /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     1492..1511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1730..1834)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(1937..2176,1..349)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
       61 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      121 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      181 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      241 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      301 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      361 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      421 gttatcccct gattctgtgg ataaccgtgc ggcccgagag catcgatcct agcatggatg
      481 catttgacag ctagctcagt cctaggtata atgctagcgg taaaattgtc aacacgcagt
      541 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
      601 caccgagtcg gtgcccggct tatcggtcag tttcacctga tttacgtaaa aacccgcttc
      661 ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtccacgac gctataccca
      721 aaagaaaaag cttgaaagaa tcgatcgatc ccgaatgagt agtacagcgg ccgcgattat
      781 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      841 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      901 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      961 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg gacccacgct
     1021 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1081 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1141 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1201 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1321 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1381 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1441 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1501 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1561 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1621 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1681 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1741 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1801 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1861 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1921 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1981 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2041 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2101 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2161 tgttaccagt ggctgc
//
