LOCUS       N-term_AAV-ABE8e-SpCas9(S55R)_(CA21)        7132 bp ds-DNA     circular SYN 05-OCT-2025
DEFINITION  derivative of JL645-Cbh_v5-N-term_AAV.
ACCESSION
VERSION     .
KEYWORDS    pX601-GFP.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct.
REFERENCE   1  (bases 1 to 8138)
  AUTHORS   Ye L, Wang J, Tan Y, Beyer AI, Xie F, Muench MO and Kan YW
  TITLE     Genome editing using CRISPR-Cas9 to create the HPFH genotype in
            HSPCs: An approach for treating sickle cell disease and
            beta-thalassemia.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Sep 20;113(38):10661-5. doi:
            10.1073/pnas.1612075113. Epub 2016 Sep 6.
  PUBMED    27601644
REFERENCE   2  (bases 1 to 8138)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 20, 2017 from SnapGene Console 4.0.0a8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     reporter        122..282
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="lacZ_a reporter"
                     /Transferred_Similarity="100.00%"
                     /label="lacZ_a reporter"
     misc_feature    300..606
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="f1 origin"
                     /Transferred_Similarity="99.67%"
                     /label="f1 origin"
                     /note="Geneious type: origin of replication"
     promoter        799..827
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="amp prom"
                     /Transferred_Similarity="100.00%"
                     /label="amp prom"
     marker          869..1729
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="amp marker"
                     /Transferred_Similarity="99.42%"
                     /label="amp marker"
     CDS             869..1729
                     /vntifkey=4
                     /label="Amp-R"
     misc_feature    1884..2503
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="pBR322 origin"
                     /Transferred_Similarity="99.68%"
                     /label="pBR322 origin"
                     /note="Geneious type: origin of replication"
     promoter        2812..2841
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="lac prom"
                     /Transferred_Similarity="100.00%"
                     /label="lac prom"
     repeat_region   2917..3046
                     /vntifkey=34
                     /modified_by="jonlevy"
                     /label="5'-ITR pSub201(+)"
     promoter        3063..3867
                     /label="CBh promoter"
     promoter        3071..3356
                     /NC_006273.2="175569-175266"
                     /created_by="User"
                     /modified_by="User"
                     /label="CMV Enhancer"
     promoter        3369..3630
                     /created_by="User"
                     /X00182.1="280-540, +2"
                     /modified_by="User"
                     /label="Chicken B-actin promoter"
     promoter        3607..3612
                     /created_by="User"
                     /label="CBA TATA Box"
     promoter        3631..3770
                     /X00182.1="541-677"
                     /created_by="User"
                     /label="Chicken B-actin intron"
     promoter        3637
                     /created_by="User"
                     /label="putative cba +1"
     promoter        3725..3733
                     /created_by="User"
                     /modified_by="User"
                     /label="b-actin SD (MaxEnt)"
     promoter        3776..3832
                     /NC_001510.1="2312-2368"
                     /created_by="User"
                     /modified_by="User"
                     /label="MVM Intron"
     promoter        3778..3786
                     /created_by="User"
                     /label="MVM SD"
     promoter        3833..3867
                     /From="methods of http://dx.doi.org/10.1038/sj.mt.6300355"
                     /created_by="User"
                     /label="MVM Intron 2"
     promoter        complement(3860..3864)
                     /created_by="User"
                     /label="MVM SA (MaxEnt)"
     motif           3874..3883
                     /created_by="benjaminkleinstiver"
                     /label="Kozak"
     CDS             3880..3936
                     /Color="#6bba70"
                     /created_by="Kleinstiver Lab 1"
                     /label="BPNLS"
     CDS             3934..4434
                     /created_by="Kleinstiver Lab 1"
                     /modified_by="bpk6"
                     /Color="#ffbbbb"
                     /label="TadA*8e"
     misc_feature    4000..4002
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="W23R"
     misc_feature    4039..4041
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="H36L"
     misc_feature    4075..4077
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="P48A"
     misc_feature    4084..4086
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="R51L"
     misc_feature    4183..4185
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="L84F"
     misc_feature    4249..4251
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="A106V"
     misc_feature    4255..4257
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="D108N"
     misc_feature    4258..4260
                     /created_by="bpk6"
                     /Color="#0000ff"
                     /modified_by="bpk6"
                     /label="A109S"
     misc_feature    4264..4266
                     /created_by="bpk6"
                     /Color="#0000ff"
                     /modified_by="bpk6"
                     /label="T111R"
     misc_feature    4288..4290
                     /created_by="bpk6"
                     /Color="#0000ff"
                     /modified_by="bpk6"
                     /label="D119N"
     misc_feature    4297..4299
                     /created_by="bpk6"
                     /Color="#0000ff"
                     /modified_by="bpk6"
                     /label="H122N"
     misc_feature    4300..4302
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="H123Y"
     misc_feature    4369..4371
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="S146C"
     misc_feature    4372..4374
                     /created_by="bpk6"
                     /Color="#0000ff"
                     /modified_by="bpk6"
                     /label="D147 (changed back from D147Y in ABE7.10)"
     misc_feature    4378..4380
                     /created_by="bpk6"
                     /Color="#0000ff"
                     /modified_by="bpk6"
                     /label="F149Y"
     misc_feature    4387..4389
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="R152P"
     misc_feature    4396..4398
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="E155V"
     misc_feature    4399..4401
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="I156F"
     misc_feature    4402..4404
                     /Color="#ff0059"
                     /created_by="bpk6"
                     /label="K157N"
     misc_feature    4429..4431
                     /created_by="bpk6"
                     /Color="#0000ff"
                     /modified_by="bpk6"
                     /label="T166I"
     misc_feature    4432..4434
                     /created_by="bpk6"
                     /Color="#0000ff"
                     /modified_by="bpk6"
                     /label="D167N"
     CDS             4435..4530
                     /Color="#ff9911"
                     /created_by="Kleinstiver Lab 1"
                     /label="linker"
     CDS             4528..>6246
                     /note="hSpCas9"
                     /modified_by="jonlevy"
                     /modified_by="ben"
                     /label="nSpCas9(D10A/S55R) until E573"
     misc_difference 4555..4557
                     /created_by="benjaminkleinstiver"
                     /label="D10A"
     misc_difference 4690..4692
                     /created_by="bpk6"
                     /modified_by="ben"
                     /label="S55R"
     CDS             6247..6552
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /Color="#3333ff"
                     /label="NpuN"
     CDS             6553..6615
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /Transferred_From="C-terminal BPNLS"
                     /Transferred_Similarity="100.00%"
                     /Color="#6bba70"
                     /label="BPNLS"
     CDS             6616..6618
                     /Color="#000000"
                     /created_by="benjaminkleinstiver"
                     /label
     terminator      6624..6784
                     /created_by="User"
                     /modified_by="User"
                     /label="WPRE_Gamma"
     Terminator      6873..7100
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="bGH_PA term"
                     /Transferred_Similarity="100.00%"
                     /label="bGH_PA term"
     repeat_region   complement(7109..106)
                     /vntifkey=34
                     /modified_by="jonlevy"
                     /label="3'-ITR pSub201(+)"
ORIGIN      
        1 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
       61 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta
      121 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      181 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      241 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
      301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      481 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      541 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      601 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
      661 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
      721 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      781 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      841 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      901 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      961 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1021 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1081 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1141 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1201 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1261 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1321 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1381 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1441 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1501 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1561 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1621 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1681 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1741 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1801 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1861 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1921 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1981 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2041 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2101 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2161 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2221 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2281 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2341 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2401 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2461 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2521 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2581 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2641 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     2701 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     2761 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     2821 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     2881 cagctatgac catgattacg ccagatttaa ttaaggctgc gcgctcgctc gctcactgag
     2941 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     3001 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg cctctagatc
     3061 agggtacccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     3121 cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag
     3181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     3241 cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta
     3301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     3361 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     3421 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc
     3481 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     3541 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     3601 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgacgct gccttcgccc
     3661 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
     3721 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag
     3781 gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct
     3841 gcctgaaatc actttttttc aggttggacc ggtgccacca tgaaacggac agccgacgga
     3901 agcgagttcg agtcaccaaa gaagaagcgg aaagtctctg aggtggagtt ttcccacgag
     3961 tactggatga gacatgccct gaccctggcc aagagggcac gcgatgagag ggaggtgcct
     4021 gtgggagccg tgctggtgct gaacaataga gtgatcggcg agggctggaa cagagccatc
     4081 ggcctgcacg acccaacagc ccatgccgaa attatggccc tgagacaggg cggcctggtc
     4141 atgcagaact acagactgat tgacgccacc ctgtacgtga cattcgagcc ttgcgtgatg
     4201 tgcgccggcg ccatgatcca ctctaggatc ggccgcgtgg tgtttggcgt gaggaacagc
     4261 aaaagaggcg ccgcaggctc cctgatgaac gtgctgaact accccggcat gaatcaccgc
     4321 gtcgaaatta ccgagggaat cctggcagat gaatgtgccg ccctgctgtg cgatttctac
     4381 cggatgccta gacaggtgtt caatgctcag aagaaggccc agagctccat caactccgga
     4441 ggatctagcg gaggctcctc tggctctgag acacctggca caagcgagag cgcaacacct
     4501 gaaagcagcg ggggcagcag cggggggtca gacaagaagt acagcatcgg cctggccatc
     4561 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     4621 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
     4681 ctgttcgaca gaggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     4741 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     4801 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     4861 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     4921 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     4981 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     5041 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     5101 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     5161 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     5221 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac
     5281 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     5341 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     5401 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     5461 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     5521 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     5581 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     5641 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     5701 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     5761 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     5821 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     5881 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     5941 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     6001 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     6061 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     6121 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     6181 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     6241 atcgagtgcc tgtcctacga gacagagatc ctgacagtgg agtatggcct gctgccaatc
     6301 ggcaagatcg tggagaagag gatcgagtgt accgtgtact ctgtggataa caatggcaac
     6361 atctatacac agcccgtggc acagtggcac gataggggag agcaggaggt gttcgagtat
     6421 tgcctggagg acggcagcct gatcagggca accaaggacc acaagttcat gacagtggat
     6481 ggccagatgc tgcccatcga cgagattttc gagcgggagc tggacctgat gagagtggat
     6541 aacctgccta atagcggagg cagtaaaaga acagcagacg ggagtgagtt tgagcccaag
     6601 aaaaagagaa aggtgtaaga tctgataatc aacctctgga ttacaaaatt tgtgaaagat
     6661 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     6721 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     6781 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6841 ctcggctgtt gggcactgac aattccgtgg tgcgactgtg ccttctagtt gccagccatc
     6901 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6961 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     7021 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     7081 ggatgcggtg ggctctatgg gcggccgcag gaacccctag tgatggagtt gg
//
