LOCUS       sequence-217170-        2637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting unit plasmid. The sgRNA targeting unit plasmids 
            contain connector ConLS, ConR1, and one sgRNA transcriptional unit. 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2637)
  AUTHORS   Geng P, Leonard SP, Mishler DM, Barrick JE
  TITLE     Synthetic Genome Defenses against Selfish DNA Elements Stabilize 
            Engineered Bacteria against Evolutionary Failure.
  JOURNAL   ACS Synth Biol. 2019 Mar 15;8(3):521-531. doi: 
            10.1021/acssynbio.8b00426. Epub 2019 Feb 15.
  PUBMED    30703321
REFERENCE   2  (bases 1 to 2637)
  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"; doi: 
            "10.1021/acssynbio.8b00426"; journalName: "ACS Synth Biol"; date: 
            "2019-03-15- 15"; volume: "8"; issue: "3"; pages: "521-531"
COMMENT     Sequence Label: pJ23119-sgRNA
FEATURES             Location/Qualifiers
     source          1..2637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        285..319
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        340..415
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(join(870..1661,1662..1730))
                     /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     1493..1512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1731..1835)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1938..2526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2427..2446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattcgcat ctagatggta gagccacaaa cagccggtac aagcaacgat ctccaggacc
       61 atctgaatca tgcgcggatg acacgaactc acgacggcga tcacagacat taacccacag
      121 tacagacact gcgacaacgt ggcaattcgt cgcaataccg tctcactgaa ctggccgata
      181 attgcagacg aacgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
      241 ccttacgcat ctgtgcggta tttcacaccg catatgctgg atccttgaca gctagctcag
      301 tcctaggtat aatactagta gtgtaattcg gggcagaatg ttttagagct agaaatagca
      361 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      421 ttgaattctc tagagtcgac ctgcagaagc ttagatctat taccctgtta tccctactcg
      481 agttcatgtg cagctccatc agcatccgta atcgttaatc cgcaaataac gtaaaaaccc
      541 gcttcggcgg gtttttttat ggggggagtt tagggaaaga gcatttgtca gctgccaatg
      601 agacgacggg gtcatcacgg ctcatcatgc gccaaacaaa tgtgtgcaat acacgctcgg
      661 atgactgcat gatgaccgca ctgactgggg acagcagatc cacctaagcc tgtgagagaa
      721 gcagacaccc gacagatcaa ggcagttaac tagtgcactg cagtacagcg gccgcgatta
      781 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      841 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      901 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      961 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc
     1021 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1081 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1141 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1201 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1261 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1321 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1381 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1441 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1501 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1561 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1621 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1681 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1741 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1801 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1861 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1921 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1981 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2041 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2101 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2161 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2221 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2281 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2341 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2401 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2461 tttcgtcacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2521 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2581 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc cgcccct
//
