LOCUS       New_DNA                 5996 bp ds-DNA     circular     03-JUN-2018
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Insert from pCR8-dCasRxHsu-28-45-18 652 to 3719
COMMENT     Backbone from pAC90-pmaxGW-D55-60,22-61-69,23-91-7 2650 to 967
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          4025..4035
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          1..947
                     /organism="pmaxCloning"
                     /mol_type="other DNA"
                     /label=source:pmaxCloning
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          4169..5996
                     /organism="pmaxCloning"
                     /mol_type="other DNA"
                     /label=source:pmaxCloning(1)
                     /ApEinfo_label=source:pmaxCloning
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             3981..4007
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
                     /label=HA
                     /ApEinfo_fwdcolor=#e9d024
                     /ApEinfo_revcolor=#e9d024
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        163..715
                     /label=CMV_immearly_promoter
                     /ApEinfo_fwdcolor=#ffaae6
                     /ApEinfo_revcolor=#ff389a
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    4026..4035
                     /label=attL2
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    218..505
                     /label=CAG_enhancer
                     /ApEinfo_fwdcolor=#33ff00
                     /ApEinfo_revcolor=#bfc4ff
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          968..991
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO(1)
                     /ApEinfo_label=source:pCR8/GW/TOPO
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        661..742
                     /label=CMV_promoter
                     /ApEinfo_fwdcolor=#ffaae6
                     /ApEinfo_revcolor=#ff389a
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    complement(968..984)
                     /label=attL1
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          960..967
                     /organism="pLenti6/Ubc/V5-DEST Gateway (ViraPower)"
                     /mol_type="other DNA"
                     /label=source:pLenti6/Ubc/V5-DEST Gateway (ViraPower)
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          4036..4156
                     /organism="pLenti6/Ubc/V5-DEST Gateway (ViraPower)"
                     /mol_type="other DNA"
                     /label=source:pLenti6/Ubc/V5-DEST Gateway (ViraPower)(1)
                     /ApEinfo_label=source:pLenti6/Ubc/V5-DEST Gateway
                     (ViraPower)
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      complement(4463..5081)
                     /label=pBR322_origin
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    960..967
                     /label=attR1
                     /ApEinfo_fwdcolor=#ffaa89
                     /ApEinfo_revcolor=#deff79
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    987..991
                     /label=EcoRIHS
                     /ApEinfo_fwdcolor=#ff8b8e
                     /ApEinfo_revcolor=#ff8b8e
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    672..692
                     /label=CMV_fwd_primer
                     /ApEinfo_fwdcolor=#33ff00
                     /ApEinfo_revcolor=#bfc4ff
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          1008..4007
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             1014..1034
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="PKKKRKV"
                     /label=SV40 NLS
                     /ApEinfo_fwdcolor=#e9d024
                     /ApEinfo_revcolor=#e9d024
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    813..945
                     /label=chim_int
                     /ApEinfo_fwdcolor=#33ff00
                     /ApEinfo_revcolor=#bfc4ff
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             3951..3971
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="PKKKRKV"
                     /label=SV40 NLS(1)
                     /ApEinfo_label=SV40 NLS
                     /ApEinfo_fwdcolor=#e9d024
                     /ApEinfo_revcolor=#e9d024
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    complement(4103..4122)
                     /label=AC_V5_R
                     /ApEinfo_fwdcolor=cyan
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             complement(5146..5991)
                     /translation="MGGDRDETNPDKCFNNIMIEQDGLHAGSPAAWVERLFGYDWAQQ
                     TIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVT
                     EAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIER
                     ARTRMEAGLVDQDDLDEEHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENG
                     RFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRL
                     LDEFF*"
                     /label=ORF frame 3
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    1008..4007
                     /label=Hsu_dCasRx
                     /ApEinfo_fwdcolor=cyan
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    complement(4170..4189)
                     /label=EBV_rev_primer
                     /ApEinfo_fwdcolor=#33ff00
                     /ApEinfo_revcolor=#bfc4ff
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     gene            complement(5149..5928)
                     /gene="NeoR/KanR (variant)"
                     /translation="MGGDRDETNPDKCFNNIMIEQDGLHAGSPAAWVERLFGYDWAQQ
                     TIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVT
                     EAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIER
                     ARTRMEAGLVDQDDLDEEHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENG
                     RFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRL
                     LDEFF*"
                     /label=NeoR/KanR (variant)
                     /ApEinfo_fwdcolor=#0004ff
                     /ApEinfo_revcolor=#ff00a8
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    4103..4144
                     /label=V5
                     /ApEinfo_fwdcolor=#ffaa89
                     /ApEinfo_revcolor=#deff79
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacagggaaa aaaCAATTGA
      961 CAAGTTTGTA CAAAAAAGCA GGCTCCGAAT TcACCGGTgc cgccaccATG AGCCCCAAGA
     1021 AGAAGAGAAA GGTGGAGGCC AGCATCGAAA AAAAAAAGTC CTTCGCCAAG GGCATGGGCG
     1081 TGAAGTCCAC ACTCGTGTCC GGCTCCAAAG TGTACATGAC AACCTTCGCC GAAGGCAGCG
     1141 ACGCCAGGCT GGAAAAGATC GTGGAGGGCG ACAGCATCAG GAGCGTGAAT GAGGGCGAGG
     1201 CCTTCAGCGC TGAAATGGCC GATAAAAACG CCGGCTATAA GATCGGCAAC GCCAAATTCA
     1261 GCCATCCTAA GGGCTACGCC GTGGTGGCTA ACAACCCTCT GTATACAGGA CCCGTCCAGC
     1321 AGGATATGCT CGGCCTGAAG GAAACTCTGG AAAAGAGGTA CTTCGGCGAG AGCGCTGATG
     1381 GCAATGACAA TATTTGTATC CAGGTGATCC ATAACATCCT GGACATTGAA AAAATCCTCG
     1441 CCGAATACAT TACCAACGCC GCCTACGCCG TCAACAATAT CTCCGGCCTG GATAAGGACA
     1501 TTATTGGATT CGGCAAGTTC TCCACAGTGT ATACCTACGA CGAATTCAAA GACCCCGAGC
     1561 ACCATAGGGC CGCTTTCAAC AATAACGATA AGCTCATCAA CGCCATCAAG GCCCAGTATG
     1621 ACGAGTTCGA CAACTTCCTC GATAACCCCA GACTCGGCTA TTTCGGCCAG GCCTTTTTCA
     1681 GCAAGGAGGG CAGAAATTAC ATCATCAATT ACGGCAACGA ATGCTATGAC ATTCTGGCCC
     1741 TCCTGAGCGG ACTGGCGCAC TGGGTGGTCG CTAACAACGA AGAAGAGTCC AGGATCTCCA
     1801 GGACCTGGCT CTACAACCTC GATAAGAACC TCGACAACGA ATACATCTCC ACCCTCAACT
     1861 ACCTCTACGA CAGGATCACC AATGAGCTGA CCAACTCCTT CTCCAAGAAC TCCGCCGCCA
     1921 ACGTGAACTA TATTGCCGAA ACTCTGGGAA TCAACCCTGC CGAATTCGCC GAACAATATT
     1981 TCAGATTCAG CATTATGAAA GAGCAGAAAA ACCTCGGATT CAATATCACC AAGCTCAGGG
     2041 AAGTGATGCT GGACAGGAAG GATATGTCCG AGATCAGGAA AAATCATAAG GTGTTCGACT
     2101 CCATCAGGAC CAAGGTCTAC ACCATGATGG ACTTTGTGAT TTATAGGTAT TACATCGAAG
     2161 AGGATGCCAA GGTGGCTGCC GCCAATAAGT CCCTCCCCGA TAATGAGAAG TCCCTGAGCG
     2221 AGAAGGATAT CTTTGTGATT AACCTGAGGG GCTCCTTCAA CGACGACCAG AAGGATGCCC
     2281 TCTACTACGA TGAAGCTAAT AGAATTTGGA GAAAGCTCGA AAATATCATG CACAACATCA
     2341 AGGAATTTAG GGGAAACAAG ACAAGAGAGT ATAAGAAGAA GGACGCCCCT AGACTGCCCA
     2401 GAATCCTGCC CGCTGGCCGT GATGTTTCCG CCTTCAGCAA ACTCATGTAT GCCCTGACCA
     2461 TGTTCCTGGA TGGCAAGGAG ATCAACGACC TCCTGACCAC CCTGATTAAT AAATTCGATA
     2521 ACATCCAGAG CTTCCTGAAG GTGATGCCTC TCATCGGAGT CAACGCTAAG TTCGTGGAGG
     2581 AATACGCCTT TTTCAAAGAC TCCGCCAAGA TCGCCGATGA GCTGAGGCTG ATCAAGTCCT
     2641 TCGCTAGAAT GGGAGAACCT ATTGCCGATG CCAGGAGGGC CATGTATATC GACGCCATCC
     2701 GTATTTTAGG AACCAACCTG TCCTATGATG AGCTCAAGGC CCTCGCCGAC ACCTTTTCCC
     2761 TGGACGAGAA CGGAAACAAG CTCAAGAAAG GCAAGCACGG CATGAGAAAT TTCATTATTA
     2821 ATAACGTGAT CAGCAATAAA AGGTTCCACT ACCTGATCAG ATACGGTGAT CCTGCCCACC
     2881 TCCATGAGAT CGCCAAAAAC GAGGCCGTGG TGAAGTTCGT GCTCGGCAGG ATCGCTGACA
     2941 TCCAGAAAAA ACAGGGCCAG AACGGCAAGA ACCAGATCGA CAGGTACTAC GAAACTTGTA
     3001 TCGGAAAGGA TAAGGGCAAG AGCGTGAGCG AAAAGGTGGA CGCTCTCACA AAGATCATCA
     3061 CCGGAATGAA CTACGACCAA TTCGACAAGA AAAGGAGCGT CATTGAGGAC ACCGGCAGGG
     3121 AAAACGCCGA GAGGGAGAAG TTTAAAAAGA TCATCAGCCT GTACCTCACC GTGATCTACC
     3181 ACATCCTCAA GAATATTGTC AATATCAACG CCAGGTACGT CATCGGATTC CATTGCGTCG
     3241 AGCGTGATGC TCAACTGTAC AAGGAGAAAG GCTACGACAT CAATCTCAAG AAACTGGAAG
     3301 AGAAGGGATT CAGCTCCGTC ACCAAGCTCT GCGCTGGCAT TGATGAAACT GCCCCCGATA
     3361 AGAGAAAGGA CGTGGAAAAG GAGATGGCTG AAAGAGCCAA GGAGAGCATT GACAGCCTCG
     3421 AGAGCGCCAA CCCCAAGCTG TATGCCAATT ACATCAAATA CAGCGACGAG AAGAAAGCCG
     3481 AGGAGTTCAC CAGGCAGATT AACAGGGAGA AGGCCAAAAC CGCCCTGAAC GCCTACCTGA
     3541 GGAACACCAA GTGGAATGTG ATCATCAGGG AGGACCTCCT GAGAATTGAC AACAAGACAT
     3601 GTACCCTGTT CGCAAACAAG GCCGTCGCCC TGGAAGTGGC CAGGTATGTC CACGCCTATA
     3661 TCAACGACAT TGCCGAGGTC AATTCCTACT TCCAACTGTA CCATTACATC ATGCAGAGAA
     3721 TTATCATGAA TGAGAGGTAC GAGAAAAGCA GCGGAAAGGT GTCCGAGTAC TTCGACGCTG
     3781 TGAATGACGA GAAGAAGTAC AACGATAGGC TCCTGAAACT GCTGTGTGTG CCTTTCGGCT
     3841 ACTGTATCCC CAGGTTTAAG AACCTGAGCA TCGAGGCCCT GTTCGATAGG AACGAGGCCG
     3901 CCAAGTTCGA CAAGGAGAAA AAGAAGGTGT CCGGCAATTC CGGATCCGGA CCTAAGAAAA
     3961 AGAGGAAGGT GGCGGCCGCT TACCCATACG ATGTTCCAGA TTACGCTtga TTAATTAAGA
     4021 ATTCGACCCA GCTTTCTTGT ACAAAGTGGT TGATATCCAG CACAGTGGCG GCCGCTCGAG
     4081 TCTAGAGGGC CCGCGGTTCG AAGGTAAGCC TATCCCTAAC CCTCTCCTCG GTCTCGATTC
     4141 TACGCGTACC GGTTAGCAAT TGttttttcg atgagtttgg acaaaccaca actagaatgc
     4201 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4261 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4321 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt acttaagagg
     4381 gggagaccaa agggcgagac gttaaggcct cacgtgacat gtgagcaaaa ggccagcaaa
     4441 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4501 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4561 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4621 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4681 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4741 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4801 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4861 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4921 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4981 cttgatccgg caaacaaacc acgctggtag cggtggtttt tttgtttgca agcagcagat
     5041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5101 tcagtggaac gaaaactcac gttaagggat tttggtcatg ccgtctcaga agaactcgtc
     5161 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     5221 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     5281 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     5341 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     5401 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     5461 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     5521 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     5581 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     5641 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     5701 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     5761 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     5821 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     5881 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     5941 aatattattg aagcatttat cagggttcgt ctcgtcccgg tctcctccca tgcatg
//
