LOCUS       sequence-214550-        2641 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 2641)
  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 2641)
  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..2641
                     /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(874..1665,1666..1734))
                     /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     1497..1516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1735..1839)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1942..2530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2431..2450
                     /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 gctggctgcc
      601 aatgagacga cggggtcatc acggctcatc atgcgccaaa caaatgtgtg caatacacgc
      661 tcggatgact gcatgatgac cgcactgact ggggacagca gatccaccta agcctgtgag
      721 agaagcagac acccgacaga tcaaggcagt taactagtgc actgcagtac agcggccgcg
      781 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      841 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      901 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      961 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc
     1021 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1081 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1141 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1201 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1261 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1321 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1381 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1441 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1501 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1561 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1621 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1681 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1741 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1801 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1861 acctgtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1921 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1981 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2041 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2101 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2161 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2221 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2281 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2341 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2401 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2461 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2521 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2581 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg cggccgcccc
     2641 t
//
