LOCUS       Exported                3368 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Plasmid designed to clone a specific spacer into BsaI sites to 
            generate a CRISPR Array.
ACCESSION   .
VERSION     .
KEYWORDS    pSEVA231-CRISPR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3368)
  AUTHORS   Velazquez E, Lorenzo V, Al-Ramahi Y
  TITLE     Recombination-Independent Genome Editing through 
            CRISPR/Cas9-Enhanced TargeTron Delivery.
  JOURNAL   ACS Synth Biol. 2019 Sep 20;8(9):2186-2193. doi: 
            10.1021/acssynbio.9b00293. Epub 2019 Sep 3.
  PUBMED    31419111
REFERENCE   2  (bases 1 to 3368)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3368
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     repeat_region   88..123
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   154..189
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_feature    complement(190..321)
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     terminator      400..494
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(615..634)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             626..1441
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            1608..1716
                     /note="incP origin of transfer"
     CDS             complement(1727..2389)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2390..3161
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      3276..3362
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 caactcaaca agtctcagtg tgctgaagtt ttgggaccat tcaaaacagc atagctctaa
      121 aacgagacct ttgagcttcc gagactggtc tcagttttgg gaccattcaa aacagcatag
      181 ctctaaaacc tcgtagacta tttttgtcta aaaaatttcg taatcgcact atttgtctca
      241 gctagacttc agtcttgaaa agcccctgta ttactgcatt tattaagagt attataccat
      301 atttttagtt attaagaaat aggatcctct agagtcgacc tgcaggcatg caagcttgcg
      361 gccgcgtcgt gactgggaaa accctggcga ctagtcttgg actcctgttg atagatccag
      421 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
      481 ttattggtga gaatccaggg gtccccaata attacgattt aaatttgtgt ctcaaaatct
      541 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
      601 cataaacagt aatacaaggg gtgttatgag ccatattcag cgtgaaacga gctgtagccg
      661 tccgcgtctg aacagcaaca tggatgcgga tctgtatggc tataaatggg cgcgtgataa
      721 cgtgggtcag agcggcgcga ccatttatcg tctgtatggc aaaccggatg cgccggaact
      781 gtttctgaaa catggcaaag gcagcgtggc gaacgatgtg accgatgaaa tggtgcgtct
      841 gaactggctg accgaattta tgccgctgcc gaccattaaa cattttattc gcaccccgga
      901 tgatgcgtgg ctgctgacca ccgcgattcc gggcaaaacc gcgtttcagg tgctggaaga
      961 atatccggat agcggcgaaa acattgtgga tgcgctggcc gtgtttctgc gtcgtctgca
     1021 tagcattccg gtgtgcaact gcccgtttaa cagcgatcgt gtgtttcgtc tggcccaggc
     1081 gcagagccgt atgaacaacg gcctggtgga tgcgagcgat tttgatgatg aacgtaacgg
     1141 ctggccggtg gaacaggtgt ggaaagaaat gcataaactg ctgccgttta gcccggatag
     1201 cgtggtgacc cacggcgatt ttagcctgga taacctgatt ttcgatgaag gcaaactgat
     1261 tggctgcatt gatgtgggcc gtgtgggcat tgcggatcgt tatcaggatc tggccattct
     1321 gtggaactgc ctgggcgaat ttagcccgag cctgcaaaaa cgtctgtttc agaaatatgg
     1381 cattgataat ccggatatga acaaactgca atttcatctg atgctggatg aatttttcta
     1441 ataattaatt ggaccgcggt ccgcgcgttg tccttttccg ctgcataacc ctgcttcggg
     1501 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
     1561 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
     1621 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
     1681 gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg ccggccctac cggcgcggca
     1741 gcgttacccg tgtcggcggc tccaacggct cgccatcgtc cagaaaacac ggctcatcgg
     1801 gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc cgtttcggtc aaggctggca
     1861 ggtctggttc catgcccgga atgccgggct ggctgggcgg ctcctcgccg gggccggtcg
     1921 gtagttgctg ctcgcccgga tacagggtcg ggatgcggcg caggtcgcca tgccccaaca
     1981 gcgattcgtc ctggtcgtcg tgatcaacca ccacggcggc actgaacacc gacaggcgca
     2041 actggtcgcg gggctggccc cacgccacgc ggtcattgac cacgtaggcc gacacggtgc
     2101 cggggccgtt gagcttcacg acggagatcc agcgctcggc caccaagtcc ttgactgcgt
     2161 attggaccgt ccgcaaagaa cgtccgatga gcttggaaag tgtcttctgg ctgaccacca
     2221 cggcgttctg gtggcccatc tgcgccacga ggtgatgcag cagcattgcc gccgtgggtt
     2281 tcctcgcaat aagcccggcc cacgcctcat gcgctttgcg ttccgtttgc acccagtgac
     2341 cgggcttgtt cttggcttga atgccgattt ctctggactg cgtggccatg cttatctcca
     2401 tgcggtaggg gtgccgcacg gttgcggcac catgcgcaat cagctgcaac ttttcggcag
     2461 cgcgacaaca attatgcgtt gcgtaaaagt ggcagtcaat tacagatttt ctttaaccta
     2521 cgcaatgagc tattgcgggg ggtgccgcaa tgagctgttg cgtacccccc ttttttaagt
     2581 tgttgatttt taagtctttc gcatttcgcc ctatatctag ttctttggtg cccaaagaag
     2641 ggcacccctg cggggttccc ccacgccttc ggcgcggctc cccctccggc aaaaagtggc
     2701 ccctccgggg cttgttgatc gactgcgcgg ccttcggcct tgcccaaggt ggcgctgccc
     2761 ccttggaacc cccgcactcg ccgccgtgag gctcgggggg caggcgggcg ggcttcgccc
     2821 ttcgactgcc cccactcgca taggcttggg tcgttccagg cgcgtcaagg ccaagccgct
     2881 gcgcggtcgc tgcgcgagcc ttgacccgcc ttccacttgg tgtccaaccg gcaagcgaag
     2941 cgcgcaggcc gcaggccgga ggcttttccc cagagaaaat taaaaaaatt gatggggcaa
     3001 ggccgcaggc cgcgcagttg gagccggtgg gtatgtggtc gaaggctggg tagccggtgg
     3061 gcaatccctg tggtcaagct cgtgggcagg cgcagcctgt ccatcagctt gtccagcagg
     3121 gttgtccacg ggccgagcga agcgagccag ccggtggccg ctcgcggcca tcgtccacat
     3181 atccacgggc tggcaaggga gcgcagcgac cgcgcagggc gaagcccgga gagcaagccc
     3241 gtaggggggg cgcgcccagc tgtctagggc ggcggatttg tcctactcag gagagcgttc
     3301 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     3361 tgatgcct
//
