LOCUS       JL645-Cbh_v5_AAV        7727 bp ds-DNA     circular SYN 28-NOV-2019
DEFINITION  .
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
     repeat_region   complement(7704..106)
                     /vntifkey=34
                     /modified_by="jonlevy"
                     /label="3'-ITR pSub201(+)"
     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        800..828
                     /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          870..1730
                     /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             870..1730
                     /vntifkey=4
                     /label="Amp-R"
     misc_feature    1885..2504
                     /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        2813..2842
                     /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   2918..3047
                     /vntifkey=34
                     /modified_by="jonlevy"
                     /label="5'-ITR pSub201(+)"
     promoter        3064..3868
                     /label="CBh promoter"
     promoter        3072..3357
                     /NC_006273.2="175569-175266"
                     /created_by="User"
                     /modified_by="User"
                     /label="CMV Enhancer"
     promoter        3370..3631
                     /created_by="User"
                     /X00182.1="280-540, +2"
                     /modified_by="User"
                     /label="Chicken B-actin promoter"
     promoter        3608..3613
                     /created_by="User"
                     /label="CBA TATA Box"
     promoter        3632..3771
                     /X00182.1="541-677"
                     /created_by="User"
                     /label="Chicken B-actin intron"
     promoter        3638
                     /created_by="User"
                     /label="putative cba +1"
     promoter        3726..3734
                     /created_by="User"
                     /modified_by="User"
                     /label="b-actin SD (MaxEnt)"
     promoter        3777..3833
                     /NC_001510.1="2312-2368"
                     /created_by="User"
                     /modified_by="User"
                     /label="MVM Intron"
     promoter        3779..3787
                     /created_by="User"
                     /label="MVM SD"
     promoter        3834..3868
                     /From="methods of http://dx.doi.org/10.1038/sj.mt.6300355"
                     /created_by="User"
                     /label="MVM Intron 2"
     promoter        complement(3861..3865)
                     /created_by="User"
                     /label="MVM SA (MaxEnt)"
     CDS             3881..3937
                     /created_by="jonlevy"
                     /Transferred_From="N-terminal BPNLS"
                     /Transferred_Similarity="100.00%"
                     /label="BPNLS"
     CDS             3938..5029
                     /created_by="jonlevy"
                     /label="ABE 6.20"
     CDS             5030..5125
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="GGS-XTEN-GGS Linker"
     CDS             5126..>6841
                     /note="hSpCas9"
                     /modified_by="jonlevy"
     CDS             6842..7147
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="NpuN"
     CDS             7148..7210
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /Transferred_From="C-terminal BPNLS"
                     /Transferred_Similarity="100.00%"
                     /label="BPNLS"
     terminator      7219..7379
                     /created_by="User"
                     /modified_by="User"
                     /label="WPRE_Gamma"
     Terminator      7468..7695
                     /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"
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 tattaacgtt tataatttca ggtggcatct ttcggggaaa tgtgcgcgga acccctattt
      781 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      841 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      901 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      961 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1021 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1081 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1141 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1201 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1261 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1321 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1381 taccaaacga cgagcgtgac accacgatgc ctgtagtaat ggtaacaacg ttgcgcaaac
     1441 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1501 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1561 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1621 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1681 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     1741 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1801 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1861 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1921 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1981 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2041 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2101 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2161 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2221 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2281 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2341 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2401 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2461 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2521 tggccttttg ctgcggtttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2581 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2641 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     2701 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     2761 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     2821 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     2881 acagctatga ccatgattac gccagattta attaaggctg cgcgctcgct cgctcactga
     2941 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     3001 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg gcctctagat
     3061 cagggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3121 ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc aatgggtgga
     3181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     3241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt acatgacctt
     3301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     3361 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     3421 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg
     3481 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
     3541 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
     3601 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc tgccttcgcc
     3661 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     3721 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag ctgagcaaga
     3781 ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac ctggagcacc
     3841 tgcctgaaat cacttttttt caggttggac cggtgccacc atgaaacgga cagccgacgg
     3901 aagcgagttc gagtcaccaa agaagaagcg gaaagtctct gaagtcgagt ttagccacga
     3961 gtattggatg aggcacgcac tgaccctggc aaagcgagca tgggatgaaa gagaagtccc
     4021 cgtgggcgcc gtgctggtgc acaacaatag agtgatcgga gagggatgga acaggccaat
     4081 cggccgccac gaccctaccg cacacgcaga gatcatggca ctgaggcagg gaggcctggt
     4141 catgcagaat taccgcctga tcgatgccac cctgtatgtg acactggagc catgcgtgat
     4201 gtgcgcagga gcaatgatcc acagcaggat cggaagagtg gtgttcggag cacgggacgc
     4261 caagaccggc gcagcaggct ccctgatgga tgtgctgcac caccccggca tgaaccaccg
     4321 ggtggagatc acagagggaa tcctggcaga cgagtgcgcc gccctgctga gcgatttctt
     4381 tagaatgcgg agacaggaga tcaaggccca gaagaaggca cagagctcca ccgactctgg
     4441 aggatctagc ggaggatcct ctggaagcga gacaccaggc acaagcgagt ccgccacacc
     4501 agagagctcc ggcggctcct ccggaggatc ctctgaggtg gagttttccc acgagtactg
     4561 gatgagacat gccctgaccc tggccaagag ggcacgcgat gagagggagg tgcctgtggg
     4621 agccgtgctg gtgctgaaca atagagtgat cggcgagggc tggaacagag ccatcggcct
     4681 gcacgaccca acagcccatg ccgaaattat ggccctgaga cagggcggcc tggtcatgca
     4741 gaactacaga ctgattgacg ccaccctgta cgtgacattc gagccttgcg tgatgtgcgc
     4801 cggcgccatg atccactcta ggatcggccg cgtggtgttt ggcgtgagga acgcaaaaac
     4861 cggcgccgca ggctccctga tggacgtgct gcactacccc ggcatgaatc accgcgtcga
     4921 aattaccgag ggaatcctgg cagatgaatg tgccgccctg ctgtgctatt tctttcggat
     4981 gcctagacag gtgttcaatg ctcagaagaa ggcccagagc tccaccgact ccggaggatc
     5041 tagcggaggc tcctctggct ctgagacacc tggcacaagc gagagcgcaa cacctgaaag
     5101 cagcgggggc agcagcgggg ggtcagacaa gaagtacagc atcggcctgg ccatcggcac
     5161 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     5221 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     5281 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     5341 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     5401 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     5461 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     5521 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     5581 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     5641 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     5701 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     5761 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     5821 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     5881 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     5941 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     6001 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     6061 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     6121 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     6181 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     6241 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     6301 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     6361 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     6421 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     6481 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     6541 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     6601 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     6661 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     6721 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     6781 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     6841 gtgcctgtcc tacgagacag agatcctgac agtggagtat ggcctgctgc caatcggcaa
     6901 gatcgtggag aagaggatcg agtgtaccgt gtactctgtg gataacaatg gcaacatcta
     6961 tacacagccc gtggcacagt ggcacgatag gggagagcag gaggtgttcg agtattgcct
     7021 ggaggacggc agcctgatca gggcaaccaa ggaccacaag ttcatgacag tggatggcca
     7081 gatgctgccc atcgacgaga ttttcgagcg ggagctggac ctgatgagag tggataacct
     7141 gcctaatagc ggaggcagta aaagaacagc agacgggagt gagtttgagc ccaagaaaaa
     7201 gagaaaggtg taagatctga taatcaacct ctggattaca aaatttgtga aagattgact
     7261 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     7321 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     7381 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     7441 ctgttgggca ctgacaattc cgtggtgcga ctgtgccttc tagttgccag ccatctgttg
     7501 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     7561 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     7621 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     7681 cggtgggctc tatgggcggc cgcaggaacc cctagtgatg gagttgg
//
