LOCUS       sequence-259037-        5254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for an easy-to-construct crRNA delivery vector using 
            hybridized oligonucleotides. Constitutive transcription of crRNA, 
            Spectinomycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5254)
  AUTHORS   Krumbach K, Sonntag CK, Eggeling L, Marienhagen J
  TITLE     CRISPR/Cas12a Mediated Genome Editing To Introduce Amino Acid 
            Substitutions into the Mechanosensitive Channel MscCG of 
            Corynebacterium glutamicum.
  JOURNAL   ACS Synth Biol. 2019 Dec 11. doi: 10.1021/acssynbio.9b00361.
  PUBMED    31790583
REFERENCE   2  (bases 1 to 5254)
  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: "ACS Synth 
            Biol. 2019 Dec 11. doi: 10.1021/acssynbio.9b00361."
COMMENT     Sequence Label: pJYScr
FEATURES             Location/Qualifiers
     source          1..5254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4125..4206
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4298..4325
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4419..4436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4590..5178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4670..4689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actaataacg taacgtgact ggcaagagat atttttaaaa caatgaatag gtttacactt
       61 actttagttt tatggaaatg aaagatcata tcatatataa tctagaataa aattaactaa
      121 aataattatt atctagataa aaaatttaga agccaatgaa atctataaat aaactaaatt
      181 aagtttattt aattaacaac tatggatata aaataggtac taatcaaaat agtgaggagg
      241 atatatttga atacatacga acaaattaat aaagtgaaaa aaatacttcg gaaacattta
      301 aaaaataacc ttattggtac ttacatgttt ggatcaggag ttgagagtgg actaaaacca
      361 aatagtgatc ttgacttttt agtcgtcgta tctgaaccat tgacagatca aagtaaagaa
      421 atacttatac aaaaaattag acctatttca aaaaaaatag gagataaaag caacttacga
      481 tatattgaat taacaattat tattcagcaa gaaatggtac cgtggaatca tcctcccaaa
      541 caagaattta tttatggaga atggttacaa gagctttatg aacaaggata cattcctcag
      601 aaggaattaa attcagattt aaccataatg ctttaccaag caaaacgaaa aaataaaaga
      661 atatacggaa attatgactt agaggaatta ctacctgata ttccattttc tgatgtgaga
      721 agagccatta tggattcgtc agaggaatta atagataatt atcaggatga tgaaaccaac
      781 tctatattaa ctttatgccg tatgatttta actatggaca cgggtaaaat cataccaaaa
      841 gatattgcgg gaaatgcagt ggctgaatct tctccattag aacataggga gagaattttg
      901 ttagcagttc gtagttatct tggagagaat attgaatgga ctaatgaaaa tgtaaattta
      961 actataaact atttaaataa cagattaaaa aaattataaa aaaattgaaa aaatggtgga
     1021 aacacttttt tcaatttttt tgttttatta tttaatattt gggaaatatt cattctaatt
     1081 ggtaatcaga ttttagaaaa caataaaccc ttgcatatga tatcgatgta cagatctttt
     1141 tttaaggcag ttattggtgc ccttggaaat gaccgaccaa gcgacgccca acctgccatc
     1201 acgagatttc gattccaccg ccgccttcta tgaaagccat caatcctgcc tatttgccac
     1261 gtttaacaag gtagttaagc gttcatttac gaagaaaaca cgataagctg cacaaatacc
     1321 tgaaaaagtt gaacgccccg tgagcgggaa ctcacagggc gtcggctaac ccccagtcat
     1381 cagctgggag aaagcactca agacatgact ctagccgatc cgcaggacac agtcacagct
     1441 agcgcgtgga aattttccgc cgatctgttc gacacccacc ccgaactagc gctgcgctca
     1501 cgcggctgga cggcagaaga tcgccgcgaa ctgctcgctc acctgggacg cgaaagcttc
     1561 cagggcagca agacaagaga tttcgcgagc gcctggatta aaaacccgga taccggcgaa
     1621 acccaaccaa agctctaccg ggctggctca aaagcgctga cgcggtgcca gtacgttgcg
     1681 ctgacgcacg cgcaacatgc cgcggtgatc gtgcttgaca tcgatgtgcc cagccaccag
     1741 gccggcggga agattgagca cgtaaacccg caggtctacg cgattttaga gaaatgggca
     1801 cgcctagaaa aagcgccggc ttggatcggc gtgaatccgc tgagcgggaa atgccagctc
     1861 atctggctca ttgacccggt gtatgccgca gcaggtaaaa ccagcccaaa tatgcgcctg
     1921 ctggctgcaa cgacggaaga aatgactcgt gttttcggcg ctgaccaggc tttttcgcat
     1981 aggctgagcc ggtggccgct gcacgtctca gacgatccga cagcctataa atggcactgc
     2041 cagcatgatc gtgtggatcg gctggccgac ctaatggaga ttgctcgaac gatgaccgga
     2101 tcacagaagc cgaaaaagta cattgagcag gacttttcca gcggacgcgc ccgcattgaa
     2161 gcggcacaac gcgccaccgc agaagccaag gcgctagcga ttttggacgc gagcctgccg
     2221 agcgccctgg acgcgtccgg cgacctgatc gacggcgtgc gagtgctctg gacaaatcca
     2281 gagcgcgcag cgcgcgacga gaccgcgttt cgccacgcgt tgaccgtggg ataccagctc
     2341 aaagctgctg gtgagcgcct aaaagatgcc aagatcatcg acgcgtatga agtggcgtac
     2401 aacgttgccc aggcggtcgg tgcagacggc cgggagccgg atcttcccgc catgcgtgat
     2461 cgcctgacga tggcgcgtcg tgtgcgcggc tacgtggcta aaggccagcc agtcgtccct
     2521 gctcgtcggg tggaaacgca gagcagccga gggcggaaag ctctagcgac gatggggcga
     2581 cggggcgcag ctacatcgaa tgcacgcaga tgggctgacc cagaaagtaa gtatgcgcag
     2641 gagacgcgac agcgattagc ggaagcaaac aaacgccgag aaatgacagg cgagttgctc
     2701 gaacttcgcg tcaaaactgc gatcctggat gcccgttctc aatcggttgc tgatccctcg
     2761 actcgtgagc ttgcaggcga actaggtgtc agtgaaaggc gcatccaaca agtcagaaag
     2821 gcacttggaa tggaagctaa acgcggccgt ccacgggctg aaaactaata aacgaaacac
     2881 cgtcagcaga aaacggttcc cccctttagg ggtcccgtcc ttgctctggc tctcacttgc
     2941 cctcaccctc cgctatccac gggctgaaaa ctaataaacg aaacaccgtc agcagaaaac
     3001 ggttcccccc ctttagggtg tctcgctcct agctctgatc cctccccggt tcctccccgg
     3061 cctgattttt aagggggctc acgctgtcgg cagagaacgg ttcccccgcc ttctgctctg
     3121 gctcttcctc gactccctcc ccctcaaaaa tctcctcgag tctagattga cagctagctc
     3181 agtcctaggt ataatggatc ccacgagtat agaacgctga gattgattcc ggccggaaag
     3241 aaaggtgatt cgcaatacaa ttatttgcga ttagcttact agcttttatt gtggctttat
     3301 ttcaacgggc tggaacgtaa tgtatgcaat ctccactaaa ttaaattaca ctctccgacc
     3361 tcgttctagt ttcacttaat atttcgccct cagcaactag agaagttaaa ggagagacga
     3421 tttgaaaata ttgtggcctt tttgaacact gtggtgaacc tttcaatatt ctccttaaag
     3481 aggcatgaag aatccacggc aaaaacttta gacgctaatt atggggcctg acccctggaa
     3541 agtagacaca ggaaatttaa tcaagaagca gatttcagga tagcagaacc ccgaatctca
     3601 tacatcgcag gtgtgagatg accacaccag gattgcctaa gactcgtgtt gtacctggta
     3661 caccacttca aatacctccc gacggcacac caactggttc gcaaacgtct tggaatcctg
     3721 cgcggaactt cccgcttgaa tgcggcatta aacgactgaa gccagcgaat tatccgcacc
     3781 gatacccaca gcacccatca actgacaaac cacccgactt tgtgcagatg tcctggaata
     3841 cttgcgaggt gtacacactg ccatgttctt cagtggaaca cagcatcctt tacgatgtct
     3901 agttggtctt tagccatcat caacgcatcc tgaaccaacg acgtcggatt aggtcttagt
     3961 tgccatgatc ggtaggttac ctttttgtcc caaattgttg ggacattagg tgtctaccaa
     4021 gcaggagcca gatccatgta tattcttcct gcacaattac agccaatagc ttacgatctt
     4081 taatgcatat acgaaggtgc aataagcgta tgatcttcga acataaaacg aaaggctcag
     4141 tcgaaagaca gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     4201 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     4261 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     4321 ctttttgcgt ttctacaaac tcttaagctt catatgcggt gtgaaatacc gcacagatgc
     4381 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5221 aacgaaaact cacgttaagg gattttggtc atga
//
