LOCUS       pgRNA                   4217 bp    DNA     circular UNA 08-JAN-2020
DEFINITION  J23119-lacZ gRNA, sacB, pBR322 ori, Cm (aka pZZ45).
ACCESSION   urn.local...t-9du8wb2
VERSION     urn.local...t-9du8wb2
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     modified_base   1..357
     primer_bind     1..25
                     /standard_name="ZZ50"
     primer_bind     290..314
                     /standard_name="ZZ31"
     promoter        358..392
     misc_feature    393..412
                     /standard_name="N20 Custom crRNA Sequence"
     misc_RNA        413..494
                     /standard_name="sgRNA"
     primer_bind     470..514
                     /standard_name="ZZ52"
     misc_feature    complement(578..602)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="ZZ58"
     misc_feature    703..1341
                     /note="Geneious type: marker"
                     /note="Tn9"
                     /product="ampicillin acetyl transferase"
                     /db_xref="GI:15554330"
                     /standard_name="cat"
     primer_bind     1065..1089
                     /standard_name="ZZ72"
     misc_feature    complement(1149..1173)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="ZZ71"
     primer_bind     1219..1243
                     /standard_name="ZZ64"
     misc_feature    1327..1366
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="ZZ363"
     CDS             complement(1415..2836)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGIS
                     HITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHG
                     YHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQT
                     QEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIF
                     DGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEES
                     LFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIAS
                     NTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLN
                     KTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLN
                     IKGKKTSVVKDSILEQGQLTVNK"
                     /standard_name="sacB"
     primer_bind     complement(3258..3297)
                     /standard_name="ZZ362"
     misc_feature    3268..3307
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="ZZ365"
     misc_feature    complement(3368..3397)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="ZZ361"
     rep_origin      3437..4056
     primer_bind     3527..3551
                     /standard_name="ZZ35"
     misc_feature    complement(3626..3650)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="ZZ34"
ORIGIN      
        1 accatggctg gtgaccacgt cgtggaatgc cttcgaattc agcacctgca catgggacgt
       61 cgacctgagg taattataac ccgggcccta tatatggatc caattgcaat gatcatcatg
      121 acagatctgc gcgcgatcga tatcagcgct ttaaatttgc gcatgctagc tatagttcta
      181 gaggtaccgg ttgttaacgt tagccggcta cgtatactcc ggaatattaa taggcctagg
      241 atgcatatgg cggccgcctg cagctggcgc catcgatacg cgtacgtcgc gaccgcggac
      301 atgtacagag ctcgagaagt actagtggcc acgtgggccg tgcaccttaa gcttggcttg
      361 acagctagct cagtcctagg tataatgcta gcnnnnnnnn nnnnnnnnnn nngttttaga
      421 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
      481 gtcggtgctt ttttggcgcg cctacctgtg acggaagatc acttcgcaga ataaataaat
      541 cctggtgtcc ctgttgatac cgggaagccc tgggccaact tttggcgaaa atgagacgtt
      601 gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
      661 ttttttgagt tgtcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
      721 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
      781 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
      841 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
      901 atgaatgctc atccggaatt acgtatggca atgaaatacg gtgagctggt gatatgggat
      961 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     1021 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     1081 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     1141 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     1201 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     1261 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag atgcttaatg
     1321 aatacaacag tactgcgatg aatgggattc acctttatgt tgataagaaa taaaagaaaa
     1381 tgccaatagg atatcggcat tttcttttgc gtttttattt gttaactgtt aattgtcctt
     1441 gttcaaggat gctgtctttg acaacagatg ttttcttgcc tttgatgttc agcaggaagc
     1501 ttggcgcaaa cgttgattgt ttgtctgcgt agaatcctct gtttgtcata tagcttgtaa
     1561 tcacgacatt gtttcctttc gcttgaggta cagcgaagtg tgagtaagta aaggttacat
     1621 cgttaggatc aagatccatt tttaacacaa ggccagtttt gttcagcggc ttgtatgggc
     1681 cagttaaaga attagaaaca taaccaagca tgtaaatatc gttagacgta atgccgtcaa
     1741 tcgtcatttt tgatccgcgg gagtcagtga acaggtacca tttgccgttc attttaaaga
     1801 cgttcgcgcg ttcaatttca tctgttactg tgttagatgc aatcagcggt ttcatcactt
     1861 ttttcagtgt gtaatcatcg tttagctcaa tcataccgag agcgccgttt gctaactcag
     1921 ccgtgcgttt tttatcgctt tgcagaagtt tttgactttc ttgacggaag aatgatgtgc
     1981 ttttgccata gtatgctttg ttaaataaag attcttcgcc ttggtagcca tcttcagttc
     2041 cagtgtttgc ttcaaatact aagtatttgt ggcctttatc ttctacgtag tgaggatctc
     2101 tcagcgtatg gttgtcgcct gagctgtagt tgccttcatc gatgaactgc tgtacatttt
     2161 gatacgtttt tccgtcaccg tcaaagattg atttataatc ctctacaccg ttgatgttca
     2221 aagagctgtc tgatgctgat acgttaactt gtgcagttgt cagtgtttgt ttgccgtaat
     2281 gtttaccgga gaaatcagtg tagaataaac ggatttttcc gtcagatgta aatgtggctg
     2341 aacctgacca ttcttgtgtt tggtctttta ggatagaatc atttgcatcg aatttgtcgc
     2401 tgtctttaaa gacgcggcca gcgtttttcc agctgtcaat agaagtttcg ccgacttttt
     2461 gatagaacat gtaaatcgat gtgtcatccg catttttagg atctccggct aatgcaaaga
     2521 cgatgtggta gccgtgatag tttgcgacag tgccgtcagc gttttgtaat ggccagctgt
     2581 cccaaacgtc caggcctttt gcagaagaga tatttttaat tgtggacgaa tcgaattcag
     2641 gaacttgata tttttcattt ttttgctgtt cagggatttg cagcatatca tggcgtgtaa
     2701 tatgggaaat gccgtatgtt tccttatatg gcttttggtt cgtttctttc gcaaacgctt
     2761 gagttgcgcc tcctgccagc agtgcggtag taaaggttaa tactgttgct tgttttgcaa
     2821 actttttgat gttcatcgtt catgtctcct tttttatgta ctgtgttagc ggtctgcttc
     2881 ttccagccct cctgtttgaa gatggcaagt tagttacgca caataaaaaa agacctaaaa
     2941 tatgtaaggg gtgacgccaa agtatacact ttgcccttta cacattttag gtcttgcctg
     3001 ctttatcagt aacaaacccg cgcgatttac ttttcgacct cattctatta gactctcgtt
     3061 tggattgcaa ctggtctatt ttcctctttt gtttgataga aaatcataaa aggatttgca
     3121 gactacgggc ctaaagaact aaaaaatcta tctgtttctt ttcattctct gtatttttta
     3181 tagtttctgt tgcatgggca taaagttgcc tttttaatca caattcagaa aatatcataa
     3241 tatctcattt cactaaataa tagtgaacgg caggtatatg tgctgtcaga ccaagtttac
     3301 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3361 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3421 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3481 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3541 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3601 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3661 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3721 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3781 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3841 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3901 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3961 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4021 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4081 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4141 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4201 tcagtgagcg aggaagc
//
