pLH02

LOCUS       pLH02                9528 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Genome editing for Lactobacillus brevis ATCC367.
ACCESSION   .
VERSION     .
KEYWORDS    pLH02
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9528)
  AUTHORS   Huang H, Song X, Yang S
  TITLE     Development of a RecE/T-Assisted CRISPR-Cas9 Toolbox for 
            Lactobacillus.
  JOURNAL   Biotechnol J. 2019 Jul;14(7):e1800690. doi: 10.1002/biot.201800690. 
            Epub 2019 May 20.
  PUBMED    30927506
REFERENCE   2  (bases 1 to 9528)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9528
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    complement(303..336)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(1348..1381)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(6941..6952)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      8654..9199
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     9100..9119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9316..9333
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgtgttgct ttgattgata gccaaaaagc agcagttgat aaagcaatta ctgatattgc
       61 tgaaaaattg taatttataa ataaaaatca ccttttagag gtggtttttt tatttataaa
      121 ttattcgttt gatttcgctt tcgatagaac aatcaaagcg agaataagga agataaatcc
      181 cataagggcg ggagcagaat gtccgagact aatgtaaatt tgtccaccaa ttaaaggacc
      241 gataacgcgc tcgagcctga tagaaacaga agccactgga gcacgtttaa acaatttaaa
      301 tctaccgttc gtataatgta tgctatacga agttatgaca atgtcttagg cgttaaggtc
      361 gttttagccg atggtcgcga agttaagtaa ggtaccatgc agtttaaatt cggtcctcgg
      421 gatatgataa gattaatagt tttagctatt aatctttttt tatttttatt taagaatggc
      481 ttaataaagc ggttactttg gatttttgtg agcttggact agaaaaaaac ttcacaaaat
      541 gctatactag gtagataaaa atttaggagg catatcaaat gaactttaat aaaattgatt
      601 tagacaattg gaagagaaaa gagatattta atcattattt gaaccaacaa acgactttta
      661 gtataaccac agaaattgat attagtgttt tataccgaaa cataaaacaa gaaggatata
      721 aattttaccc tgcatttatt ttcttagtga caagggtgat aaactcaaat acagctttta
      781 gaactggtta caatagcgac ggagagttag gttattggga taagttagag ccactttata
      841 caatttttga tggtgtatct aaaacattct ctggtatttg gactcctgta aagaatgact
      901 tcaaagagtt ttatgattta tacctttctg atgtagagaa atataatggt tcggggaaat
      961 tgtttcccaa aacacctata cctgaaaatg ctttttctct ttctattatt ccatggactt
     1021 catttactgg gtttaactta aatatcaata ataatagtaa ttaccttcta cccattatta
     1081 cagcaggaaa attcattaat aaaggtaatt caatatattt accgctatct ttacaggtac
     1141 atcattctgt ttgtgatggt tatcatgcag gattgtttat gaactctatt caggaattgt
     1201 cagataggcc taatgactgg cttttataat atgagataat gccgactgta cttggatcct
     1261 aaacgcaatt gatgattggt tcggaaggca cgttaggaat cattaccgaa gtaatcgtta
     1321 aactgttgcc gattccgcta gggacccata acttcgtata atgtatgcta tacgaacggt
     1381 acagaccggg gtcgaccctt caatagagtt cttaacgtta atccgaaaaa aactaacgtt
     1441 aatattaaaa aataagatcc gcttgtgaat tatgtataat ttgattagac taaagaatag
     1501 gagaaagtat gatgatattt aaaaaacttt ctcgttaaga taggttgttg gtgagcatgt
     1561 tatatacgga tgtatcggtt tccttaatgc aaaattttgt tgctatctta ttaatttttc
     1621 tattatatag atatattcaa agaaagataa catttaaacg gatcatatta gatattttaa
     1681 tagcgattat tttttcaata ttatatctgt ttatttcaga tgcgtcatta cttgtaatgg
     1741 tattaatgcg attagggtgg cattttcatc aacaaaaaga aaataagata aaaacgactg
     1801 atacagctaa tttaattcta attatcgtga tccagttatt gttagttgcg gttgggacta
     1861 ttattagtca gtttaccata tcgattatca aaagtgattt cagccaaaat atattgaaca
     1921 atagtgcaac agatataact ttattaggta ttttctttgc tgttttattt gacggcttgt
     1981 tctttatatt attgaagaat aagcggactg aattacaaca tttaaatcaa gaaatcattg
     2041 aattttcgtt agaaaaacaa tattttatat ttatatttat tttatttata gtaatagaaa
     2101 ttattttagc agttgggaat cttcaaggag taacagccac gatattatta accattatca
     2161 ttattttttg tgtccttatc gggatgactt tttggcaagt gatgcttttt ttgaaggctt
     2221 attcgattcg ccaagaagcc aatgaccaat tggtccggaa tcaacaactt caagattatc
     2281 tagtcaatat cgaacagcag tacaccgaat tacggcgatt taagcatgat tatcaaaaca
     2341 tcttattatc gttggagagt tttgccgaaa agggcgatca gcaacagttt aaggcgtatt
     2401 accaagaatt attagcacaa cggccaattc aaagtgaaat ccaaggggca gtcattgcac
     2461 aactcgacta cttgaaaaat gatcctattc gaggattagt cattcaaaag tttttggcag
     2521 ccaaacaggc tggtgttact ttaaaattcg aaatgaccga accaatcgaa ttagcaaccg
     2581 ctaatctatt aacggttatt cggattatcg gtattttatt agacaatgcg attgaacaag
     2641 ccgttcaaga aaccgatcaa ttggtgagtt gtgctttctt acaatctgat ggtttaatcg
     2701 aaattacgat tgaaaatacg gccagtcaag ttaagaatct ccaagcattt tcagagttag
     2761 gctattcaac gaaaggcgct ggtcggggga ctggtttagc taatgtgcag gatttgattg
     2821 ccaaacaaac caatttattc ttagaaacac agattgaaaa tagaaagtta cgacagacat
     2881 tgatgattac ggaggaaact taatttgtat cccgtttatt tattagagga tgatttacag
     2941 caacaagcga tttatcagca aattatcgcg aatacgatta tgattaacga atttgcaatg
     3001 actttaacat gcgctgccag tgatactgag acattgttgg cggcaattaa ggatcagcaa
     3061 cgaggtttat tctttttgga tatggaaatt gaggataacc gccaagccgg tttagaagtg
     3121 gcaactaaga ttcggcagat gatgccgttt gcgcaaattg tcttcattac aacccacgag
     3181 gaactgacat tattaacgtt agaacgaaaa atagcgcctt tagattacat tctcaaggac
     3241 caaacaatgg ctgaaatcaa aaggcaattg attgatgatc tattgttagc tgagaagcaa
     3301 aacgaggcgg cagcgtatca ccgagaaaat ttatttagtt ataaaatagg tcctcgcttt
     3361 ttctcattac cattaaagga agttgtttat ttatatactg aaaaagaaaa tccgggtcat
     3421 attaatttgt tagccgttac cagaaaggtt acttttccag gaaatttaaa tgcgctggaa
     3481 gcccaatatc caatgctctt tcggtgtgat aaaagttact tagttaacct atctaatatt
     3541 gccaattatg acagtaaaac acggagttta aaatttgtag atggcagtga ggcaaaagtc
     3601 tcgttccgga aatcacggga actagtggcc aaattaaaac aaatgatgta gcgcctgcag
     3661 cacgccaaat gatcccagta aaaagccacc cgcatggcgg gtggcttttt attagcccta
     3721 gaagggcttc ccacacgcat ttcagcgcct tagtgcctta gtttgtgaat cataggtggt
     3781 atagtcccga aatacccgtc taaggaattg tcagataggc ctaatgactg gcttttataa
     3841 tatgagataa tgccgactgt actttttaca gtcggttttc taatgtcact aacctgcccc
     3901 gttagttgaa gaaggttttt atattacagc tccagatcta ccggtgggcc catattaacg
     3961 tttaaccgat aaagttgaac gttaatattt tttttgcgca gaaatggtaa attgaagcat
     4021 aatagtcttg taaggtattt agctggctgg cgtaaagtat gctttataaa ataatatata
     4081 ggagtatgat tcccatgtat attacagctc cagatctacc ggtgggccca tattaacgtt
     4141 taaccgataa agttgaacgt taatattttt tttgcgcaga aatggtaaat tgaagcataa
     4201 tagtcttgta aggtatttag ctggctggcg taaagtatgc tttataaaat aatatatagg
     4261 agtatgattc ccatgcatat taacgtttaa ccgataaagt tgaacgttaa tatttttttt
     4321 gcgcagaaat ggtaaattga agcataatag tcttgtaagg tatttagctg gctggcgtaa
     4381 agtatgcttt ataaaataat atataggagt atgattccca tggacgtagt caaggtacac
     4441 acaagcagtc gtttccagcc taaagcagtg gttgctacta atttagctga actagatgac
     4501 attaaatctg atctgttcaa ggaagttcaa ttgttggccg agaatgatcg acttagcaat
     4561 gatgagattg atcggctata cagcatcagc gacgagcttg ttgcttggtc acccaatttg
     4621 gaggaggaag aataaatggc taatgaagta gcagcaacac aacgctcatt agatgcagat
     4681 gtgcaagatt caattaatca aatgatgaac caagagaacg gattgaagtt gccagcaaac
     4741 tatgccgtag gaaacgccct caagtcggca ttctttgcac taaagggaaa taacgatggg
     4801 gacttgattc aagtggctgc tcacatgcct gaaatgaaga cttcaatcgc caatgccctc
     4861 atggatatgg tggttcaagg attaacaccg gccaagaccc aagtgtactt tatcagatat
     4921 gacaatcagg ttaaaatgca gcgttcctac tttgggacac aggcggcgct aaagcgacta
     4981 tctgaggttc acgactgctg ggcaaatgta gttcatgaag gtgacggact agaaatcggt
     5041 gccgaagatg atcgtttggt cgtaagggat tggaagccga cgttggaggg actcgataag
     5101 gaaattaagt acgtttatgc cgtcattgag atggctgacg gaactcacca gcatactatc
     5161 atgaccttca aacaaatcaa aaacagctgg tcacagacac ggtctaaagg ggctgtgcag
     5221 aacaagttca gtgatgagat ggccaaacgg acggtgctta accgggccgc taagaacatt
     5281 ttaaacactt ctgatgattc agatttggtt gttggagcca tcaacaatac tacttccaac
     5341 gagtatgacg atgatcaagc agccaaagat gtgacgccta agaaggttac tgatttgatc
     5401 ggtaatgcgg acacagagga accaacgcct cctgaatcta ctgaacaaac agaaacagtt
     5461 gaaccagcag aacaagaaga accggttgat acaaatacgg acgaaagtag ctctggcgag
     5521 attcgttcta tcgaaaatat gaagccgggt gaaaagcaag acaaggacac tgtcaacgat
     5581 attttggatg gccttgaaga atctgaaaat cataaggggg atggcgacga tgcaagcagc
     5641 accgaagaag gacagggtga actcttccca ccagacgttc attctaaatt ctgataatta
     5701 ttacagtcag aaggccaata gaaacttcat gagtccaact tggtttaaga agttcgttgc
     5761 ctgtgaggca gaggcattag ctgaattgag aggtgagtgg gcacccgatg aggataagac
     5821 cgctttatta gttggtaatg acctgcacag ctattttgag tcgtctgagg cccatgaacg
     5881 gttcttagat gccaacaagg aagtgatgtt atccagtcgt ggtaagacta aaggccagct
     5941 aaagagtgag tacaagcaga ccgacatcat gattgactcg ctcaaaaacg acaagacgtt
     6001 cacacagtta tatcaaggtg aaaaggaatc aatcgttact ggcgtaattt ctggcgttaa
     6061 atggatgggc aagttggact gtttgaactt agatcgtggg tatttcatcg atctaaagac
     6121 aacacaggaa ctgtctaagc ggttttggga tagccgtacc cgtcagtgga taccattcgt
     6181 tctgaaatat gactaccagc ttcaaatggc tgtctatcgc gagctagtca agcagcagta
     6241 cggaatcgag tgtgaaccct acattgttgc agtaacaaag caaagtccgc cggataaggc
     6301 ggttatcacg attcctcatg agtatatgga ggatgccttg caacgaatag acgagcagct
     6361 accccatttt gaggatgtaa ttgctggtgt acaggcccct gttccatgtg gtagctgtgc
     6421 ttattgccgt gaacataagc agttggaaaa catcatcagc gttgatgact tactagatag
     6481 ttaggaattc ggtaccccgg gttcgaaggc gccaagcttc aaattacagc acgtgttgct
     6541 ttgattgata gccaaaaagc agcagttgat aaagcaatta ctgatattgc tgaaaaattg
     6601 taatttataa ataaaaatca ccttttagag gtggtttttt tatttataaa ttattcgttt
     6661 gatttcgctt tcgatagaac aatcaaagcg agaataagga agataaatcc cataagggcg
     6721 ggagcagaat gtccgagact aattcatgga tcgatttttt attaaaacgt ctcaaaatcg
     6781 tttctgagac gttttagcgt ttatttcgtt tagttatcgg cataatcgtt aaaacaggcg
     6841 ttatcgtagc gtaaaagccc ttgagcgtag cgtggctttg cagcgaagat gttgtctgtt
     6901 agattatgaa agccgatgac tgaatgaaat aataagcgca gcgtccttct atttcggttg
     6961 gaggaggctc aagggagttt gagggaatga aattccctca tgggtttgat tttaaaaatt
     7021 gcttgcaatt ttgccgagcg gtagcgctgg aaaatttttg aaaaaaattt ggaatttgga
     7081 aaaaaatggg gggaaaggaa gcgaattttg cttccgtact acgacccccc attaagtgcc
     7141 accccccatt aagtgccgag tgccaatttt tgtgccaaaa acgctctatc ccaactggct
     7201 caagggtttg aggggttttt caatcgccaa cgaatcgcca acgttttcgc caacgttttt
     7261 tataaatcta tatttaagta gctttattgt tgtttttatg attacaaagt gatacactaa
     7321 ttttataaaa ttatttgatt ggagtttttt aaatggtgat ttcagaatcg aaaaaaagag
     7381 ttatgatttc tctgacaaaa gagcaagata aaaaattaac agatatggcg aaacaaaaag
     7441 gtttttcaaa atctgcggtt gcggcgttag ctatagaaga atatgcaaga aaggaatcag
     7501 aacaaaaaaa ataagcgaaa gctcgcgttt ttagaaggat acgagttttc gctacttgtt
     7561 tttgataagg taattatatc atggctatta aaaatactaa agctagaaat tttggatttt
     7621 tattatatcc tgactcaatt cctaatgatt ggaaagaaaa attagagagt ttgggcgtat
     7681 ctatggctgt cagtccttta cacgatatgg acgaaaaaaa agataaagat acatggaata
     7741 gtagtgatgt tatacgaaat ggaaagcact ataaaaaacc acactatcac gttatatata
     7801 ttgcacgaaa tcctgtaaca atagaaagcg ttaggaacaa gattaagcga aaattgggga
     7861 atagttcagt tgctcatgtt gagatacttg attatatcaa aggttcatat gaatatttga
     7921 ctcatgaatc aaaggacgct attgctaaga ataaacatat atacgacaaa aaagatattt
     7981 tgaacattaa tgattttgat attgaccgct atataacact tgatgaaagc caaaaaagag
     8041 aattgaagaa tttactttta gatatagtgg atgactataa tttggtaaat acaaaagatt
     8101 taatggcttt tattcgcctt aggggagcgg agtttggaat tttaaatacg aatgatgtaa
     8161 aagatattgt ttcaacaaac tctagcgcct ttagattatg gtttgagggc aattatcagt
     8221 gtggatatag agcaagttat gcaaaggttc ttgatgctga aacgggggaa ataaaatgac
     8281 aaacaaagaa aaagagttat ttgctgaaaa tgaggaatta aaaaaagaaa ttaaggactt
     8341 aaaagagcgt attgaaagat acagagaaat ggaagttgaa ttaagtacaa caatagattt
     8401 attgagagga gggattattg aataaataaa agcccccctg acgaaagtcg cactgattgg
     8461 tgtatcattt cgtttttctt tttgtgcgga cctgcagatg cgatatcatg cgcatgcaag
     8521 cttatcgatg ataagctgtc aaacatgaga attacaactt atatcgtatg gggctgactt
     8581 caggtgctac atttgaagag ataaattgca ctgaaatcta gaaatatttt atctgattaa
     8641 taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa
     8701 aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag
     8761 gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg
     8821 cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct
     8881 tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8941 gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg
     9001 agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg
     9061 aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta
     9121 tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg
     9181 ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc
     9241 ttcctggcat cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt
     9301 cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata
     9361 ttctgctgac gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac
     9421 cctcatcagt gccaacatag taagccagta tacactccgc tagcgctgat gtccggcggt
     9481 gcttttgccg ttacgcacca ccccgtcagt agctgaacag gagggaca
//