LOCUS       JL628-Cbh_v5_AAV        7660 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(7637..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..4042
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="NpuC"
     CDS             <4043..6427
                     /codon_start=1
                     /note="hSpCas9"
                     /modified_by="jonlevy"
     CDS             6428..6718
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="genscript UGI"
     CDS             6719..6769
                     /created_by="jonlevy"
                     /label="BPNLS"
     terminator      6778..6938
                     /created_by="User"
                     /modified_by="User"
                     /label="WPRE_Gamma"
     Terminator      7027..7254
                     /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(7261..7267)
                     /created_by="User"
                     /label="U6 terminator"
     ncRNA           complement(7268..7343)
                     /created_by="User"
                     /label="RNA scaffold"
     promoter        complement(7372..7628)
                     /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 gaaagtcatc aagattgcta cacggaaata
     3961 cctgggaaag cagaacgtgt acgacatcgg cgtggagcgg gatcacaact tcgccctgaa
     4021 gaatggcttt atcgccagca attgcttcga ctccgtggaa atctccggcg tggaagatcg
     4081 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     4141 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     4201 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     4261 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     4321 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     4381 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     4441 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     4501 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     4561 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     4621 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     4681 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     4741 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     4801 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     4861 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     4921 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     4981 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     5041 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     5101 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     5161 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     5221 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     5281 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     5341 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     5401 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     5461 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     5521 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     5581 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     5641 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     5701 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     5761 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     5821 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     5881 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     5941 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     6001 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     6061 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     6121 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     6181 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     6241 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     6301 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     6361 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     6421 aggtgacagc ggcgggagcg gcgggagcgg ggggagcact aatctgagcg acatcattga
     6481 gaaggagact gggaaacagc tggtcattca ggagtccatc ctgatgctgc ctgaggaggt
     6541 ggaggaagtg atcggcaaca agccagagtc tgacatcctg gtgcacaccg cctacgacga
     6601 gtccacagat gagaatgtga tgctgctgac ctctgacgcc cccgagtata agccttgggc
     6661 cctggtcatc caggattcta acggcgagaa taagatcaag atgctgagcg gaggatccaa
     6721 aagaaccgcc gacggcagcg aattcgagcc caagaagaag aggaaagtct aagatctgat
     6781 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     6841 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     6901 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     6961 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     7021 gtggtgcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     7081 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     7141 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     7201 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggctcgag
     7261 aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc cttattttaa
     7321 cttgctattt ctagctctaa aaccgagacg cattaatcgt ctcgggtgtt tcgtcctttc
     7381 cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag
     7441 tccattttaa aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt
     7501 cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattctaat tatctctcta
     7561 acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcttcctgc
     7621 ccgaccttgc ggccgcagga acccctagtg atggagttgg
//
