LOCUS       pCas9_antiBFP           9326 bp ds-DNA     circular     24-APR-2024
DEFINITION  .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported using the Genbank importer. File name:
            addgene-plasmid-42876-sequence-212940.gbk Bacterial expression of
            Cas9 nuclease, tracrRNA and crRNA guide.
FEATURES             Location/Qualifiers
     primer          1..14
                     /label="pCas9 Forward Primer (6/1/2021)"
                     /note="sequence: gtgtcattccgctg"
     promoter        509..537
                     /label="tet promoter"
     misc_RNA        complement(855..933)
                     /label="tracrRNA"
     primer          1229..1251
                     /label="pCas9F"
                     /note="sequence: atggataagaaatactcaatagg"
     primer          1229..1251
                     /label="Cas9 portion of pCas9 forward primer (6/1/2021)"
                     /note="sequence: atggataagaaatactcaatagg"
     CDS             1229..5335
                     /label="Cas9"
     primer          complement(1711..1728)
                     /label="pCas9R_received"
                     /note="sequence: tgaccacgaaacttaatc"
     primer          5226..5245
                     /label="Guide RNA Sequencing Primer"
                     /note="sequence: gtaaacgatatacgtctaca"
     primer          complement(5321..5335)
                     /label="Cas9 portion of pCas9 reverse primer (6/1/2021)"
                     /note="sequence: tcagtcacctcctag"
     primer          complement(5321..5335)
                     /label="pCas9R"
                     /note="sequence: tcagtcacctcctag"
     misc_feature    5356..5487
                     /label="crRNA leader"
     repeat_region   5488..5523
                     /label="DR"
     repeat_region   5554..5589
                     /label="DR"
     primer_bind     complement(6018..6037)
                     /label="pBRrevBam"
     CDS             complement(7890..8549)
                     /label="CmR"
     primer_bind     8464..8483
                     /label="CAT-R"
     promoter        complement(8550..8652)
                     /label="cat promoter"
     primer_bind     complement(9044..9061)
                     /label="L4440"
     rep_origin      complement(9178..397)
                     /label="p15A ori"
     primer_bind     complement(9258..9277)
                     /label="pBR322ori-F"
     primer          complement(9312..9326)
                     /label="pCas9 Reverse Primer to amplify all of pCas9 (6/1/2021)"
                     /note="sequence: cggtaaaccgaaagg"
ORIGIN
        1 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
       61 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
      121 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
      181 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
      241 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
      301 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
      361 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
      421 aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
      481 catacgatat aagttgtaat tctcatgttt gacagcttat catcgataag ctttaatgcg
      541 gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg
      601 cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc
      661 cggtactgcc gggcctcttg cgggattacg aaatcatcct gtggagctta gtaggtttag
      721 caagatggca gcgcctaaat gtagaatgat aaaaggatta agagattaat ttccctaaaa
      781 atgataaaac aagcgttttg aaagcgcttg tttttttggt ttgcagtcag agtagaatag
      841 aagtatcaaa aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt
      901 attttaactt gctatgctgt tttgaatggt tccaacaaga ttattttata acttttataa
      961 caaataatca aggagaaatt caaagaaatt tatcagccat aaaacaatac ttaatactat
     1021 agaatgataa caaaataaac tactttttaa aagaattttg tgttataatc tatttattat
     1081 taagtattgg gtaatatttt ttgaagagat attttgaaaa agaaaaatta aagcatatta
     1141 aactaatttc ggaggtcatt aaaactatta ttgaaatcat caaactcatt atggatttaa
     1201 tttaaacttt ttattttagg aggcaaaaat ggataagaaa tactcaatag gcttagatat
     1261 cggcacaaat agcgtcggat gggcggtgat cactgatgaa tataaggttc cgtctaaaaa
     1321 gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa aaaaatctta taggggctct
     1381 tttatttgac agtggagaga cagcggaagc gactcgtctc aaacggacag ctcgtagaag
     1441 gtatacacgt cggaagaatc gtatttgtta tctacaggag attttttcaa atgagatggc
     1501 gaaagtagat gatagtttct ttcatcgact tgaagagtct tttttggtgg aagaagacaa
     1561 gaagcatgaa cgtcatccta tttttggaaa tatagtagat gaagttgctt atcatgagaa
     1621 atatccaact atctatcatc tgcgaaaaaa attggtagat tctactgata aagcggattt
     1681 gcgcttaatc tatttggcct tagcgcatat gattaagttt cgtggtcatt ttttgattga
     1741 gggagattta aatcctgata atagtgatgt ggacaaacta tttatccagt tggtacaaac
     1801 ctacaatcaa ttatttgaag aaaaccctat taacgcaagt ggagtagatg ctaaagcgat
     1861 tctttctgca cgattgagta aatcaagacg attagaaaat ctcattgctc agctccccgg
     1921 tgagaagaaa aatggcttat ttgggaatct cattgctttg tcattgggtt tgacccctaa
     1981 ttttaaatca aattttgatt tggcagaaga tgctaaatta cagctttcaa aagatactta
     2041 cgatgatgat ttagataatt tattggcgca aattggagat caatatgctg atttgttttt
     2101 ggcagctaag aatttatcag atgctatttt actttcagat atcctaagag taaatactga
     2161 aataactaag gctcccctat cagcttcaat gattaaacgc tacgatgaac atcatcaaga
     2221 cttgactctt ttaaaagctt tagttcgaca acaacttcca gaaaagtata aagaaatctt
     2281 ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat gggggagcta gccaagaaga
     2341 attttataaa tttatcaaac caattttaga aaaaatggat ggtactgagg aattattggt
     2401 gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc tttgacaacg gctctattcc
     2461 ccatcaaatt cacttgggtg agctgcatgc tattttgaga agacaagaag acttttatcc
     2521 atttttaaaa gacaatcgtg agaagattga aaaaatcttg acttttcgaa ttccttatta
     2581 tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg atgactcgga agtctgaaga
     2641 aacaattacc ccatggaatt ttgaagaagt tgtcgataaa ggtgcttcag ctcaatcatt
     2701 tattgaacgc atgacaaact ttgataaaaa tcttccaaat gaaaaagtac taccaaaaca
     2761 tagtttgctt tatgagtatt ttacggttta taacgaattg acaaaggtca aatatgttac
     2821 tgaaggaatg cgaaaaccag catttctttc aggtgaacag aagaaagcca ttgttgattt
     2881 actcttcaaa acaaatcgaa aagtaaccgt taagcaatta aaagaagatt atttcaaaaa
     2941 aatagaatgt tttgatagtg ttgaaatttc aggagttgaa gatagattta atgcttcatt
     3001 aggtacctac catgatttgc taaaaattat taaagataaa gattttttgg ataatgaaga
     3061 aaatgaagat atcttagagg atattgtttt aacattgacc ttatttgaag atagggagat
     3121 gattgaggaa agacttaaaa catatgctca cctctttgat gataaggtga tgaaacagct
     3181 taaacgtcgc cgttatactg gttggggacg tttgtctcga aaattgatta atggtattag
     3241 ggataagcaa tctggcaaaa caatattaga ttttttgaaa tcagatggtt ttgccaatcg
     3301 caattttatg cagctgatcc atgatgatag tttgacattt aaagaagaca ttcaaaaagc
     3361 acaagtgtct ggacaaggcg atagtttaca tgaacatatt gcaaatttag ctggtagccc
     3421 tgctattaaa aaaggtattt tacagactgt aaaagttgtt gatgaattgg tcaaagtaat
     3481 ggggcggcat aagccagaaa atatcgttat tgaaatggca cgtgaaaatc agacaactca
     3541 aaagggccag aaaaattcgc gagagcgtat gaaacgaatc gaagaaggta tcaaagaatt
     3601 aggaagtcag attcttaaag agcatcctgt tgaaaatact caattgcaaa atgaaaagct
     3661 ctatctctat tatctccaaa atggaagaga catgtatgtg gaccaagaat tagatattaa
     3721 tcgtttaagt gattatgatg tcgatcacat tgttccacaa agtttcctta aagacgattc
     3781 aatagacaat aaggtcttaa cgcgttctga taaaaatcgt ggtaaatcgg ataacgttcc
     3841 aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga caacttctaa acgccaagtt
     3901 aatcactcaa cgtaagtttg ataatttaac gaaagctgaa cgtggaggtt tgagtgaact
     3961 tgataaagct ggttttatca aacgccaatt ggttgaaact cgccaaatca ctaagcatgt
     4021 ggcacaaatt ttggatagtc gcatgaatac taaatacgat gaaaatgata aacttattcg
     4081 agaggttaaa gtgattacct taaaatctaa attagtttct gacttccgaa aagatttcca
     4141 attctataaa gtacgtgaga ttaacaatta ccatcatgcc catgatgcgt atctaaatgc
     4201 cgtcgttgga actgctttga ttaagaaata tccaaaactt gaatcggagt ttgtctatgg
     4261 tgattataaa gtttatgatg ttcgtaaaat gattgctaag tctgagcaag aaataggcaa
     4321 agcaaccgca aaatatttct tttactctaa tatcatgaac ttcttcaaaa cagaaattac
     4381 acttgcaaat ggagagattc gcaaacgccc tctaatcgaa actaatgggg aaactggaga
     4441 aattgtctgg gataaagggc gagattttgc cacagtgcgc aaagtattgt ccatgcccca
     4501 agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga ttctccaagg agtcaatttt
     4561 accaaaaaga aattcggaca agcttattgc tcgtaaaaaa gactgggatc caaaaaaata
     4621 tggtggtttt gatagtccaa cggtagctta ttcagtccta gtggttgcta aggtggaaaa
     4681 agggaaatcg aagaagttaa aatccgttaa agagttacta gggatcacaa ttatggaaag
     4741 aagttccttt gaaaaaaatc cgattgactt tttagaagct aaaggatata aggaagttaa
     4801 aaaagactta atcattaaac tacctaaata tagtcttttt gagttagaaa acggtcgtaa
     4861 acggatgctg gctagtgccg gagaattaca aaaaggaaat gagctggctc tgccaagcaa
     4921 atatgtgaat tttttatatt tagctagtca ttatgaaaag ttgaagggta gtccagaaga
     4981 taacgaacaa aaacaattgt ttgtggagca gcataagcat tatttagatg agattattga
     5041 gcaaatcagt gaattttcta agcgtgttat tttagcagat gccaatttag ataaagttct
     5101 tagtgcatat aacaaacata gagacaaacc aatacgtgaa caagcagaaa atattattca
     5161 tttatttacg ttgacgaatc ttggagctcc cgctgctttt aaatattttg atacaacaat
     5221 tgattgtaaa cgatatacgt ctacaaaaga agttttagat gccactctta tccatcaatc
     5281 catcactggt ctttatgaaa cacgcattga tttgagtcag ctaggaggtg actgaagtat
     5341 attttagatg aagattattt cttaataact aaaaatatgg tataatactc ttaataaatg
     5401 cagtaataca ggggcttttc aagactgaag tctagctgag acaaatagtg cgattacgaa
     5461 attttttaga caaaaatagt ctacgaggtt ttagagctat gctgttttga atggtcccaa
     5521 aacACGAGGG CACCCAGACC ATGAGAATCA AGGgttttag agctatgctg ttttgaatgg
     5581 tcccaaaact tcagcacact gagacttgtt gagttccatg ttttagagct atgctgtttt
     5641 gaatggactc cattcaacat tgccgatgat aacttgagaa agagggttaa taccagcagt
     5701 cggatacctt cctattcttt ctgttaaagc gttttcatgt tataataggc aaaagaagag
     5761 tagtgtgatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
     5821 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
     5881 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
     5941 cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     6001 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     6061 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     6121 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
     6181 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat
     6241 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     6301 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     6361 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     6421 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     6481 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
     6541 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     6601 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     6661 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcattggacc
     6721 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
     6781 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     6841 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
     6901 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
     6961 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg
     7021 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     7081 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     7141 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     7201 taaagtctgg aaacgcggaa gtcccctacg tgctgctgaa gttgcccgca acagagagtg
     7261 gaaccaaccg gtgataccac gatactatga ctgagagtca acgccatgag cggcctcatt
     7321 tcttattctg agttacaaca gtccgcaccg ctgccggtag ctccttccgg tgggcgcggg
     7381 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     7441 tgccggcagc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     7501 aagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     7561 gaacacctac atctgtatta acgaagcgct aaccgttttt atcaggctct gggaggcaga
     7621 ataaatgatc atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca
     7681 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     7741 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct
     7801 ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt
     7861 taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac
     7921 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     7981 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     8041 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     8101 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     8161 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     8221 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     8281 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga
     8341 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     8401 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     8461 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     8521 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat
     8581 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     8641 acgtgccgat caacgtctca ttttcgccaa aagttggccc ggggcttccc ggtatcaaca
     8701 gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg
     8761 caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt gtttttgagg
     8821 tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg acggggtggt
     8881 gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact ggcttactat
     8941 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     9001 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     9061 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     9121 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     9181 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     9241 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     9301 tcctgttcct gcctttcggt ttaccg
//