LOCUS       sequence-306018-        4214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Double-plasmid-based CRISPR-Cas9 system in Corynebacterium 
            glutamicum; rep oriVC. glutamicum; pMB1 oriVE. Coli; Pj23119-crRNA 
            targeting crtYf; Spcr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4214)
  AUTHORS   Zhang J, Qian F, Dong F, Wang Q, Yang J, Jiang Y, Yang S
  TITLE     De Novo Engineering of Corynebacterium glutamicum for l-Proline 
            Production.
  JOURNAL   ACS Synth Biol. 2020 Jul 17;9(7):1897-1906. doi: 
            10.1021/acssynbio.0c00249. Epub 2020 Jul 6.
  PUBMED    32627539
REFERENCE   2  (bases 1 to 4214)
  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.0c00249"; journalName: "ACS Synth Biol"; date: 
            "2020-07-17- 17"; volume: "9"; issue: "7"; pages: "1897-1906"
COMMENT     Sequence Label: pCgsgRNA_crtYf
FEATURES             Location/Qualifiers
     source          1..4214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        2259..2334
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2416..2433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2587..3175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2667..2686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ataaactatt taaataacag attaaaaaaa ttataaaaaa attgaaaaaa tggtggaaac
       61 acttttttca atttttttgt tttattattt aatatttggg aaatattcat tctaattggt
      121 aatcagattt tagaaaacaa taaacccttg catatgatat cgatgtacag atcttttttt
      181 aaggcagtta ttggtgccct tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
      241 agatttcgat tccaccgccg ccttctatga aagccatcaa tcctgcctat ttgccacgtt
      301 taacaaggta gttaagcgtt catttacgaa gaaaacacga taagctgcac aaatacctga
      361 aaaagttgaa cgccccgtga gcgggaactc acagggcgtc ggctaacccc cagtcatcag
      421 ctgggagaaa gcactcaaga catgactcta gccgatccgc aggacacagt cacagctagc
      481 gcgtggaaat tttccgccga tctgttcgac acccaccccg aactagcgct gcgctcacgc
      541 ggctggacgg cagaagatcg ccgcgaactg ctcgctcacc tgggacgcga aagcttccag
      601 ggcagcaaga caagagattt cgcgagcgcc tggattaaaa acccggatac cggcgaaacc
      661 caaccaaagc tctaccgggc tggctcaaaa gcgctgacgc ggtgccagta cgttgcgctg
      721 acgcacgcgc aacatgccgc ggtgatcgtg cttgacatcg atgtgcccag ccaccaggcc
      781 ggcgggaaga ttgagcacgt aaacccgcag gtctacgcga ttttagagaa atgggcacgc
      841 ctagaaaaag cgccggcttg gatcggcgtg aatccgctga gcgggaaatg ccagctcatc
      901 tggctcattg acccggtgta tgccgcagca ggtaaaacca gcccaaatat gcgcctgctg
      961 gctgcaacga cggaagaaat gactcgtgtt ttcggcgctg accaggcttt ttcgcatagg
     1021 ctgagccggt ggccgctgca cgtctcagac gatccgacag cctataaatg gcactgccag
     1081 catgatcgtg tggatcggct ggccgaccta atggagattg ctcgaacgat gaccggatca
     1141 cagaagccga aaaagtacat tgagcaggac ttttccagcg gacgcgcccg cattgaagcg
     1201 gcacaacgcg ccaccgcaga agccaaggcg ctagcgattt tggacgcgag cctgccgagc
     1261 gccctggacg cgtccggcga cctgatcgac ggcgtgcgag tgctctggac aaatccagag
     1321 cgcgcagcgc gcgacgagac cgcgtttcgc cacgcgttga ccgtgggata ccagctcaaa
     1381 gctgctggtg agcgcctaaa agatgccaag atcatcgacg cgtatgaagt ggcgtacaac
     1441 gttgcccagg cggtcggtgc agacggccgg gagccggatc ttcccgccat gcgtgatcgc
     1501 ctgacgatgg cgcgtcgtgt gcgcggctac gtggctaaag gccagccagt cgtccctgct
     1561 cgtcgggtgg aaacgcagag cagccgaggg cggaaagctc tagcgacgat ggggcgacgg
     1621 ggcgcagcta catcgaatgc acgcagatgg gctgacccag aaagtaagta tgcgcaggag
     1681 acgcgacagc gattagcgga agcaaacaaa cgccgagaaa tgacaggcga gttgctcgaa
     1741 cttcgcgtca aaactgcgat cctggatgcc cgttctcaat cggttgctga tccctcgact
     1801 cgtgagcttg caggcgaact aggtgtcagt gaaaggcgca tccaacaagt cagaaaggca
     1861 cttggaatgg aagctaaacg cggccgtcca cgggctgaaa actaataaac gaaacaccgt
     1921 cagcagaaaa cggttccccc ctttaggggt cccgtccttg ctctggctct cacttgccct
     1981 caccctccgc tatccacggg ctgaaaacta ataaacgaaa caccgtcagc agaaaacggt
     2041 tccccccctt tagggtgtct cgctcctagc tctgatccct ccccggttcc tccccggcct
     2101 gatttttaag ggggctcacg ctgtcggcag agaacggttc ccccgccttc tgctctggct
     2161 cttcctcgac tccctccccc tcaaaaatct cctcgagtct agattgacag ctagctcagt
     2221 cctaggtata atggatccct gcgtttaaac atatttccgt tttagagcta gaaatagcaa
     2281 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     2341 taagcttcat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     2401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2521 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2581 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2641 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2701 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2761 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2821 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2881 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2941 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3001 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3061 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3121 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3181 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3241 tttggtcatg aactaataac gtaacgtgac tggcaagaga tatttttaaa acaatgaata
     3301 ggtttacact tactttagtt ttatggaaat gaaagatcat atcatatata atctagaata
     3361 aaattaacta aaataattat tatctagata aaaaatttag aagccaatga aatctataaa
     3421 taaactaaat taagtttatt taattaacaa ctatggatat aaaataggta ctaatcaaaa
     3481 tagtgaggag gatatatttg aatacatacg aacaaattaa taaagtgaaa aaaatacttc
     3541 ggaaacattt aaaaaataac cttattggta cttacatgtt tggatcagga gttgagagtg
     3601 gactaaaacc aaatagtgat cttgactttt tagtcgtcgt atctgaacca ttgacagatc
     3661 aaagtaaaga aatacttata caaaaaatta gacctatttc aaaaaaaata ggagataaaa
     3721 gcaacttacg atatattgaa ttaacaatta ttattcagca agaaatggta ccgtggaatc
     3781 atcctcccaa acaagaattt atttatggag aatggttaca agagctttat gaacaaggat
     3841 acattcctca gaaggaatta aattcagatt taaccataat gctttaccaa gcaaaacgaa
     3901 aaaataaaag aatatacgga aattatgact tagaggaatt actacctgat attccatttt
     3961 ctgatgtgag aagagccatt atggattcgt cagaggaatt aatagataat tatcaggatg
     4021 atgaaaccaa ctctatatta actttatgcc gtatgatttt aactatggac acgggtaaaa
     4081 tcataccaaa agatattgcg ggaaatgcag tggctgaatc ttctccatta gaacataggg
     4141 agagaatttt gttagcagtt cgtagttatc ttggagagaa tattgaatgg actaatgaaa
     4201 atgtaaattt aact
//
