LOCUS       JL626-Cbh_v5_AAV        7693 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(7670..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"
     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..4621
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="rAPOBEC1"
     CDS             4622..4717
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="GGS-XTEN-GGS Linker"
     misc_feature    4640..4645
                     /Recognition_pattern="G^GATCC"
                     /Methylation="Dam:cut, Dcm:cut, CpG:cut"
                     /annotation_group="GGATCC"
                     /label="BamHI"
                     /note="Geneious type: restriction site"
     CDS             4718..6433
                     /codon_start=1
                     /note="hSpCas9"
                     /modified_by="jonlevy"
     misc_difference 4742..4744
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="D10A"
     CDS             6434..6739
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="NpuN"
     CDS             6740..6802
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /Transferred_From="C-terminal BPNLS"
                     /Transferred_Similarity="100.00%"
                     /label="BPNLS"
     terminator      6811..6971
                     /created_by="User"
                     /modified_by="User"
                     /label="WPRE_Gamma"
     Terminator      7060..7287
                     /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"
     terminator      complement(7294..7300)
                     /created_by="User"
                     /label="U6 terminator"
     ncRNA           complement(7301..7376)
                     /created_by="User"
                     /label="RNA scaffold"
     promoter        complement(7405..7661)
                     /created_by="User"
                     /label="hU6"
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 gaaagtctcc tcagagactg ggcctgtcgc
     3961 cgtcgatcca accctgcgcc gccggattga acctcacgag tttgaagtgt tctttgaccc
     4021 ccgggagctg agaaaggaga catgcctgct gtacgagatc aactggggag gcaggcactc
     4081 catctggagg cacacctctc agaacacaaa taagcacgtg gaggtgaact tcatcgagaa
     4141 gtttaccaca gagcggtact tctgccccaa taccagatgt agcatcacat ggtttctgag
     4201 ctggtcccct tgcggagagt gtagcagggc catcaccgag ttcctgtcca gatatccaca
     4261 cgtgacactg tttatctaca tcgccaggct gtatcaccac gcagacccaa ggaataggca
     4321 gggcctgcgc gatctgatca gctccggcgt gaccatccag atcatgacag agcaggagtc
     4381 cggctactgc tggcggaact tcgtgaatta ttctcctagc aacgaggccc actggcctag
     4441 gtacccacac ctgtgggtgc gcctgtacgt gctggagctg tattgcatca tcctgggcct
     4501 gcccccttgt ctgaatatcc tgcggagaaa gcagccccag ctgaccttct ttacaatcgc
     4561 cctgcagtct tgtcactatc agaggctgcc accccacatc ctgtgggcca caggcctgaa
     4621 gtctggagga tctagcggag gatcctctgg cagcgagaca ccaggaacaa gcgagtcagc
     4681 aacaccagag agcagtggcg gcagcagcgg cggcagcgac aagaagtaca gcatcggcct
     4741 ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     4801 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     4861 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     4921 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     4981 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     5041 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     5101 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     5161 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     5221 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     5281 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     5341 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     5401 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     5461 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     5521 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     5581 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     5641 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     5701 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     5761 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     5821 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     5881 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     5941 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     6001 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     6061 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     6121 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     6181 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     6241 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     6301 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     6361 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     6421 caagaaaatc gagtgcctgt cctacgagac agagatcctg acagtggagt atggcctgct
     6481 gccaatcggc aagatcgtgg agaagaggat cgagtgtacc gtgtactctg tggataacaa
     6541 tggcaacatc tatacacagc ccgtggcaca gtggcacgat aggggagagc aggaggtgtt
     6601 cgagtattgc ctggaggacg gcagcctgat cagggcaacc aaggaccaca agttcatgac
     6661 agtggatggc cagatgctgc ccatcgacga gattttcgag cgggagctgg acctgatgag
     6721 agtggataac ctgcctaata gcggaggcag taaaagaaca gcagacggga gtgagtttga
     6781 gcccaagaaa aagagaaagg tgtaagatct gataatcaac ctctggatta caaaatttgt
     6841 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     6901 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     6961 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     7021 acaggggctc ggctgttggg cactgacaat tccgtggtgc gactgtgcct tctagttgcc
     7081 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     7141 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     7201 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     7261 atgctgggga tgcggtgggc tctatggctc gagaaaaaaa gcaccgactc ggtgccactt
     7321 tttcaagttg ataacggact agccttattt taacttgcta tttctagctc taaaaccgag
     7381 acgcattaat cgtctcgggt gtttcgtcct ttccacaaga tatataaagc caagaaatcg
     7441 aaatactttc aagttacggt aagcatatga tagtccattt taaaacataa ttttaaaact
     7501 gcaaactacc caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg
     7561 tttacagtca aattaattct aattatctct ctaacagcct tgtatcgtat atgcaaatat
     7621 gaaggaatca tgggaaatag gccctcttcc tgcccgacct tgcggccgca ggaaccccta
     7681 gtgatggagt tgg
//
