LOCUS       pLVX-hyg-sgRNA1 (Linear)   8191 bp DNA     linear   SYN 13-SEP-2021
COMMENT     Created by Takara Bio USA, Inc. http://www.takarabio.com
COMMENT     Cat. Nos. 632632, 632629, 632630
FEATURES             Location/Qualifiers
     misc_feature    2..93
                     /label="sgRNA Scaffold"
                     /note="improved guide RNA scaffold for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     promoter        127..626
                     /label=PGK promoter
     CDS             644..1669
                     /label="HygR"
                     /note="confers resistance to hygromycin"
     misc_feature    1696..2284
                     /label="WPRE"
     LTR             2491..3124
                     /label="3' LTR"
     rep_origin      complement(3654..4242)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4413..5273)
                     /label="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(5274..5378)
                     /label=AmpR promoter
     polyA_signal    5426..5560
                     /label="SV40 poly(A) signal"
     LTR             5728..6361
                     /label="5' LTR"
     misc_feature    6408..6533
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7030..7263
                     /label="RRE"
     misc_feature    7755..7870
                     /label="cPPT/CTS"
     promoter        7927..8167
                     /label="U6 promoter"
ORIGIN
        1 gtttaagagc tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac
       61 ttgaaaaagt ggcaccgagt cggtgctttt tttgtgagaa ttcatctaga acgcgtgaat
      121 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
      181 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
      241 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
      301 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
      361 tagcacgtgt cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
      421 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
      481 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
      541 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
      601 cctcttcctc atctccgggc ctttcgacct gcatcccgcc accatgaaaa agcctgaact
      661 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtgt ccgacctgat
      721 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
      781 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
      841 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
      901 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
      961 cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga
     1021 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     1081 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     1141 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     1201 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     1261 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     1321 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     1381 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat
     1441 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     1501 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     1561 tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc
     1621 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagt ctagagggcc
     1681 cgacgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1741 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1801 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     1861 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     1921 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     1981 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2041 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     2101 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2161 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2221 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2281 ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca
     2341 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     2401 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     2461 ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac
     2521 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
     2581 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
     2641 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
     2701 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
     2761 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
     2821 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
     2881 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
     2941 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     3001 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     3061 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     3121 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     3181 tcagagagtg agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg
     3241 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3301 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3361 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3421 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3481 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3541 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3601 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3661 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3721 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3781 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3841 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3901 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3961 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4021 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4081 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4141 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4201 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4261 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4321 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4381 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4441 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4501 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4561 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4621 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4681 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4741 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4801 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4861 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4921 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4981 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5041 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5101 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5161 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5221 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5281 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5341 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5401 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
     5461 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5521 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
     5581 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
     5641 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
     5701 ttaaatatgt actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag
     5761 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     5821 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     5881 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     5941 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     6001 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     6061 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     6121 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     6181 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6241 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6301 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6361 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     6421 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     6481 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     6541 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     6601 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     6661 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     6721 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     6781 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     6841 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
     6901 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     6961 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     7021 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     7081 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     7141 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     7201 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     7261 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     7321 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     7381 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     7441 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     7501 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     7561 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     7621 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     7681 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     7741 tcgacggtat cgcctttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     7801 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     7861 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttatcga tagatctggg
     7921 caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     7981 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     8041 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     8101 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     8161 aaaggacgaa acaccggaga cggacgtctc t
//
