LOCUS       pFH68                  10835 bp    DNA     circular UNA 30-APR-2019
DEFINITION  Level1 ZmUBIp::SpCas9-NG (human optimized):NOSt (Position 2).
ACCESSION   urn.local...5p-aesjz9n
VERSION     urn.local...5p-aesjz9n
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     misc_feature    6..11
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     promoter        34..928
                     /label="UBIQUITIN upstream Promoter region (Zea mays)"
     5'_UTR          929..1010
                     /modified_by="hahnf"
                     /label="UBIQUITIN Untranslated Exon 1 /5' UTR (Zea mays)"
     5'_UTR          929
                     /label="Start of transcription"
     misc._feature   1009..1012
                     /label="donor splice"
     intron          1011..2020
                     /label="UBIQUITIN intron (Zea mays)"
     misc._feature   2018..2021
                     /label="acceptor splice"
     5'_UTR          2021..2022
                     /label="UBIQUITIN Untranslated Exon 2/5' UTR (Zea mays)"
     CDS             2023..6222
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="SpCas9-NG (human optimized)"
     sig_peptide     2023..2073
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="SV40 NLS"
     Misc._Feature   2023..2025
                     /label="native start of translation"
     sig_peptide     6175..6222
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="Nucleoplasmin NLS"
     Stop_codon      6223..6225
                     /created_by="hahnf"
                     /label="Stop"
     terminator      6230..6492
                     /vntifkey=43
                     /label="NOSt"
     misc_feature    complement(6503..6508)
                     /Recognition_pattern="GAAGAC(2/6)"
                     /annotation_group="GAAGAC"
                     /label="BpiI"
                     /note="Geneious type: restriction site"
     Misc._feature   complement(6524..6651)
                     /label="RB short"
     rep_origin      complement(8187..8801)
                     /modified_by="hahnf"
                     /Transferred_From="pFH13, pFH14, pFH15, pFH16, pFH17,
                     pFH18 , pFH19, pFH20, pFH21, pFH22, pFH32, pFH45, pFH46,
                     pFH47, pFH48, pFH55 , pFH76, pFH77, pFH78, pFH79, pFH91 ,
                     pFH92"
                     /Transferred_Similarity="99.84%"
                     /label="rep (pMB1)"
     CDS             complement(8964..9821)
                     /label="AmpR"
     Misc._feature   10666..10813
                     /label="LB"
ORIGIN      
        1 tctgtgaaga caagcaagaa ttcaagcttg gaggtgcagc gtgacccggt cgtgcccctc
       61 tctagagata atgagcattg catgtctaag ttataaaaaa ttaccacata ttttttttgt
      121 cacacttgtt tgaagtgcag tttatctatc tttatacata tatttaaact ttactctacg
      181 aataatataa tctatagtac tacaataata tcagtgtttt agagaatcat ataaatgaac
      241 agttagacat ggtctaaagg acaattgagt attttgacaa caggactcta cagttttatc
      301 tttttagtgt gcatgtgttc tccttttttt ttgcaaatag cttcacctat ataatacttc
      361 atccatttta ttagtacatc catttagggt ttagggttaa tggtttttat agactaattt
      421 ttttagtaca tctattttat tctattttag cctctaaatt aagaaaacta aaactctatt
      481 ttagtttttt tatttaataa tttagatata aaatagaata aaataaagtg actaaaaatt
      541 aaacaaatac cctttaagaa attaaaaaaa ctaaggaaac atttttcttg tttcgagtag
      601 ataatgccag cctgttaaac gccgtcgacg agtctaacgg acaccaacca gcgaaccagc
      661 agcgtcgcgt cgggccaagc gaagcagacg gcacggcatc tctgtcgctg cctctggacc
      721 cctctcgaga gttccgctcc accgttggac ttgctccgct gtcggcatcc agaaattgcg
      781 tggcggagcg gcagacgtga gccggcacgg caggcggcct cctcctcctc tcacggcacg
      841 gcagctacgg gggattcctt tcccaccgct ccttcgcttt cccttcctcg cccgccgtaa
      901 taaatagaca ccccctccac accctctttc cccaacctcg tgttgttcgg agcgcacaca
      961 cacacaacca gatctccccc aaatccaccc gtcggcacct ccgcttcaag gtacgccgct
     1021 cgtcctcccc ccccccccct ctctaccttc tctagatcgg cgttccggtc catggttagg
     1081 gcccggtagt tctacttctg ttcatgtttg tgttagatcc gtgtttgtgt tagatccgtg
     1141 ctgctagcgt tcgtacacgg atgcgacctg tacgtcagac acgttctgat tgctaacttg
     1201 ccagtgtttc tctttgggga atcctgggat ggctctagcc gttccgcaga cgggatcgat
     1261 ttcatgattt tttttgtttc gttgcatagg gtttggtttg cccttttcct ttatttcaat
     1321 atatgccgtg cacttgtttg tcgggtcatc ttttcatgct tttttttgtc ttggttgtga
     1381 tgatgtggtc tggttgggcg gtcgttctag atcggagtag aattctgttt caaactacct
     1441 ggtggattta ttaattttgg atctgtatgt gtgtgccata catattcata gttacgaatt
     1501 gaagatgatg gatggaaata tcgatctagg ataggtatac atgttgatgc gggttttact
     1561 gatgcatata cagagatgct ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg
     1621 gtcgttcatt cgttctagat cggagtagaa tactgtttca aactacctgg tgtatttatt
     1681 aattttggaa ctgtatgtgt gtgtcataca tcttcatagt tacgagttta agatggatgg
     1741 aaatatcgat ctaggatagg tatacatgtt gatgtgggtt ttactgatgc atatacatga
     1801 tggcatatgc agcatctatt catatgctct aaccttgagt acctatctat tataataaac
     1861 aagtatgttt tataattatt ttgatcttga tatacttgga tgatggcata tgcagcagct
     1921 atatgtggat ttttttagcc ctgccttcat acgctattta tttgcttggt actgtttctt
     1981 ttgtcgatgc tcaccctgtt gtttggtgtt acttctgcag gaatggcccc aaagaagaag
     2041 cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac
     2101 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     2161 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     2221 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     2281 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     2341 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     2401 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     2461 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     2521 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     2581 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     2641 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     2701 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     2761 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     2821 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     2881 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     2941 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     3001 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     3061 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     3121 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     3181 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     3241 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     3301 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     3361 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     3421 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     3481 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     3541 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     3601 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     3661 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     3721 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     3781 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     3841 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     3901 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     3961 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     4021 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     4081 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     4141 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     4201 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     4261 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     4321 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     4381 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     4441 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     4501 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     4561 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     4621 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     4681 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     4741 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     4801 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     4861 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     4921 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     4981 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     5041 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     5101 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     5161 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     5221 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     5281 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     5341 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     5401 cggcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     5461 tacggcggct tcgttagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     5521 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     5581 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     5641 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     5701 aagagaatgc tggcctctgc ccggttcctg cagaagggaa acgaactggc cctgccctcc
     5761 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     5821 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     5881 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     5941 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     6001 cacctgttta ccctgaccaa tctgggagcc cctcgggcct tcaagtactt tgacaccacc
     6061 atcgaccgga aggtgtaccg gagcaccaaa gaggtgctgg acgccaccct gatccaccag
     6121 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg
     6181 ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agtaagcttg tcaagcagat
     6241 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     6301 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     6361 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     6421 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     6481 ttactagatc gacgctacta ttgtcttctg cacgaagtgg tttaaactat cagtgtttga
     6541 caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt
     6601 taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg
     6661 ttccccagat caggcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta
     6721 gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg
     6781 caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga
     6841 tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa
     6901 atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt
     6961 cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg
     7021 ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag
     7081 gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc
     7141 ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta
     7201 gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg
     7261 ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc
     7321 cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg
     7381 cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca
     7441 cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag
     7501 caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg
     7561 cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc
     7621 ctcctgttca agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg
     7681 agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac
     7741 ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc
     7801 ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg
     7861 attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc
     7921 ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc
     7981 ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac
     8041 gaataccagc gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca
     8101 cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta
     8161 ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8221 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8281 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8341 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8401 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8461 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8521 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8581 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8641 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8701 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8761 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8821 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8881 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8941 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     9001 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9061 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct
     9121 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     9181 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9241 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9301 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9361 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9421 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9481 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9541 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9601 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9661 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9721 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9781 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     9841 tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga atgtatttag
     9901 aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt tttggggtga
     9961 ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc
    10021 gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg
    10081 cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt
    10141 tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac
    10201 agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc
    10261 aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca
    10321 cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg
    10381 ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct
    10441 gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca
    10501 gtgagggcca agttttccgc gaggtatcca caacgccggc ggccgcggtg tctcgcacac
    10561 ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag cccagcggcg
    10621 agggcaacca gcccggtgag cgtcgcaaag gagatcctga tctgactgat gggctgcctg
    10681 tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg gctggtggca
    10741 ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt
    10801 ttttaatgta ctggggtgga tgcagtgggc cccac
//
