LOCUS       CRISPR/Cas9-PX458-Ef1a 10213 bp ds-DNA     circular     28-JAN-2020
DEFINITION  .
ACCESSION   pVec1                                                              
VERSION     pVec1.1                                                            
FEATURES             Location/Qualifiers
     misc_feature    8533..9390
                     /label="AmpR"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    268..343
                     /label="chimeric guide RNA scaffold"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     rep_origin      7708..8014
                     /label="f1 Origin"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    6054..6059
                     /label="EcoRI"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    1854..1904
                     /label="NLS"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    6522..7226
                     /label="mCherry"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    7476..7616
                     /label="R-ITR"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     promoter        440..1773
                     /label="EF1a Promoter"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    1785..1853
                     /label="3xFLAG"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    7227..7232
                     /label="EcoRI"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    1905..6005
                     /label="hSpCsn1"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    6060..6122
                     /label="T2A"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    344..349
                     /label="U6 terminator"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        1..249
                     /label="Human U6 Promoter"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    6947..6952
                     /label="BBSI"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    6006..6053
                     /label="NLS"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    6123..6521
                     /label="H2B"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     rep_origin      9541..10208
                     /label="pUC ori"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    7236..7467
                     /label="bGH polyA"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          6505..6541
                     /label="Cherry-F"
                     /note="sequence: caccggtcgccaccatggtgagcaagggcgaggagga"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(6502..6538)
                     /label="H2B-R"
                     /note="sequence: tcctcgcccttgctcaccatggtggcgaccggtggat"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctct agaggtacca gtaattcata caaaaggact cgcccctgcc ttggggaatc
      481 ccagggaccg tcgttaaact cccactaacg tagaacccag agatcgctgc gttcccgccc
      541 cctcacccgc ccgctctcgt catcactgag gtggagaaga gcatgcgtga ggctccggtg
      601 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      661 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      721 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      781 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      841 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacgc
      901 ccctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      961 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcttggg
     1021 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
     1081 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     1141 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     1201 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     1261 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     1321 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     1381 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     1441 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1501 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1561 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1621 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1681 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1741 caaagttttt ttcttccatt tcaggtgtcg tgaccggtgc caccatggac tataaggacc
     1801 acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc
     1861 caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca
     1921 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     1981 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     2041 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     2101 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     2161 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     2221 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     2281 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     2341 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     2401 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     2461 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     2521 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     2581 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
     2641 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     2701 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     2761 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     2821 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     2881 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     2941 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     3001 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     3061 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     3121 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     3181 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     3241 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     3301 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     3361 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     3421 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     3481 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     3541 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     3601 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     3661 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     3721 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     3781 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     3841 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     3901 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     3961 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     4021 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     4081 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     4141 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     4201 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     4261 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     4321 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     4381 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     4441 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     4501 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     4561 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     4621 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     4681 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     4741 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     4801 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     4861 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     4921 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     4981 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     5041 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     5101 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     5161 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     5221 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     5281 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     5341 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     5401 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     5461 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     5521 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     5581 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     5641 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     5701 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     5761 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     5821 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     5881 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     5941 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     6001 gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggaattcg
     6061 gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
     6121 caATGCCTGA ACCCTCTAAG TCTGCTCCAG CCCCTAAAAA GGGTTCTAAG AAGGCTATCA
     6181 CTAAGGCGCA GAAGAAGGAT GGTAAGAAGC GTAAGCGCAG CCGCAAGGAG AGCTATTCTA
     6241 TCTATGTGTA CAAGGTTCTG AAGCAGGTCC ACCCCGACAC CGGCATCTCA TCCAAGGCCA
     6301 TGGGGATCAT GAACTCCTTC GTCAACGACA TCTTCGAGCG CATCGCGGGC GAGGCTTCTC
     6361 GCCTGGCTCA CTACAATAAG CGCTCGACCA TCACCTCCAG GGAGATTCAG ACGGCTGTGC
     6421 GCCTGCTGCT GCCTGGGGAG CTGGCTAAGC ATGCTGTGTC CGAGGGCACT AAGGCAGTTA
     6481 CCAAGTACAC TAGCTCtaag gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg
     6541 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     6601 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     6661 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     6721 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     6781 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     6841 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     6901 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaaa accatgggct
     6961 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     7021 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     7081 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     7141 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     7201 ccggcggcat ggacgagctg tacaaggaat tctaactaga gctcgctgat cagcctcgac
     7261 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     7321 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     7381 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     7441 ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc
     7501 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     7561 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     7621 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     7681 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7741 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     7801 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7861 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     7921 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7981 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     8041 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     8101 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     8161 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     8221 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     8281 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     8341 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     8401 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     8461 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     8521 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     8581 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     8641 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     8701 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     8761 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     8821 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     8881 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     8941 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     9001 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     9061 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     9121 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     9181 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     9241 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     9301 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     9361 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     9421 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     9481 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     9541 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     9601 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     9661 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     9721 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     9781 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     9841 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     9901 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     9961 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
    10021 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
    10081 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    10141 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
    10201 tttgctcaca tgt
//