LOCUS       JL842_Cbh_v5_AAV        7584 bp ds-DNA     circular SYN 30-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(7561..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..4621
                     /created_by="jonlevy"
                     /X-Transferred_From="JLXX-pCMV_rAPOBEC1(genscript)_Cas9n_(
                     GGS)3UGI"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="JL545-pCMV_BPNLS_rAPOBEC1(genscript)_dl
                     BE4_NpuN1"
                     /Transferred_Similarity="100.00%"
                     /label="Genscript rAPOBEC1"
     CDS             4622..4717
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="GGS-XTEN-GGS Linker"
     CDS             4718..>6316
                     /created_by="jonlevy"
                     /label="saCas9"
     CDS             6317..6622
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="NpuN"
     CDS             6623..6685
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /Transferred_From="C-terminal BPNLS"
                     /Transferred_Similarity="100.00%"
                     /label="BPNLS"
     terminator      6694..6854
                     /created_by="User"
                     /modified_by="User"
                     /label="WPRE_Gamma"
     Terminator      6943..7170
                     /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"
     misc_feature    7171..7560
                     /Original_Bases="GGTGTTTCGTCCTTTCCACAAGATATATAAAGCCAAGAAAT
                     CGAAATACTTTCAAGTTACGGTAAGCATATGATAGTCCATTTTAAAACATAATTTTAA
                     AACTGCAAACTACCCAAGAAATTATTACTTTCTACGTCACGTATTTTGTACTAATATC
                     TTTGTGTTTACAGTCAAATTAATTCTAATTATCTCTCTAACAGCCTTGTATCGTATAT
                     GCAAATATGAAGGAATCATGGGAAATAGGCCCTCTTCCTGCCCGACCTT"
                     /label="GGTGTTTCGTCCTTTCCACAAGATATATAAAGCCAAGAAATCGAAATACT
                     TTCAAGTTACGGTAAGCATATGATAGTCCATTTTAAAACATAATTTTAAAACTGCAAA
                     CTACCCAAGAAATTATTACTTTCTACGTCACGTATTTTGTACTAATATCTTTGTGTTT
                     ACAGTCAAATTAATTCTAATTATCTCTCTAACAGCCTTGTATCGTATATGCAAATATG
                     AAGGAATCATGGGAAATAGGCCCTCTTCCTGCCCGACCTT"
                     /note="Geneious type: Editing History Replacement"
     misc_RNA        complement(7177..7267)
                     /created_by="User"
                     /modified_by="User"
                     /label="Sa-sgRNA(84)"
     promoter        complement(7289..>7552)
                     /SerialCloner_Color="&h0019C0"
                     /SerialCloner_Show="True"
                     /SerialCloner_Protect="True"
                     /SerialCloner_Arrow="True"
                     /modified_by="User"
                     /label="U6 promoter"
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 cggcagcggg aagcgaaatt acattctggg
     4741 gctggccatt ggcattacat cagtgggcta tggcatcatt gactacgaga caagggacgt
     4801 gatcgacgcc ggcgtgagac tgttcaagga ggccaacgtg gagaacaatg agggccggag
     4861 atccaagagg ggagcaaggc gcctgaagcg gagaaggcgc cacagaatcc agagagtgaa
     4921 gaagctgctg ttcgattaca acctgctgac cgaccactcc gagctgtctg gcatcaatcc
     4981 ttatgaggcc agagtgaagg gcctgtccca gaagctgtct gaggaggagt ttagcgccgc
     5041 cctgctgcac ctggcaaaga ggagaggcgt gcacaacgtg aatgaggtgg aggaggacac
     5101 cggcaacgag ctgtccacaa aggagcagat cagccgcaat tccaaggccc tggaggagaa
     5161 gtatgtggcc gagctgcagc tggagcggct gaagaaggat ggcgaggtga ggggctccat
     5221 caatcgcttc aagacctctg actacgtgaa ggaggccaag cagctgctga aggtgcagaa
     5281 ggcctaccac cagctggatc agtcctttat cgatacatat atcgacctgc tggagacaag
     5341 gcgcacatac tatgagggac caggagaggg ctctcccttc ggctggaagg acatcaagga
     5401 gtggtacgag atgctgatgg gccactgcac ctattttcca gaggagctga gaagcgtgaa
     5461 gtacgcctat aacgccgatc tgtacaacgc cctgaatgac ctgaacaacc tggtcatcac
     5521 cagggatgag aacgagaagc tggagtacta tgagaagttc cagatcatcg agaacgtgtt
     5581 caagcagaag aagaagccta cactgaagca gatcgccaag gagatcctgg tgaacgagga
     5641 ggacatcaag ggctaccgcg tgacctccac aggcaagcca gagttcacca atctgaaggt
     5701 gtatcacgat atcaaggaca tcacagcccg gaaggagatc atcgagaacg ccgagctgct
     5761 ggatcagatc gccaagatcc tgaccatcta tcagagctcc gaggacatcc aggaggagct
     5821 gaccaacctg aatagcgagc tgacacagga ggagatcgag cagatcagca atctgaaggg
     5881 ctacaccggc acacacaacc tgagcctgaa ggccatcaat ctgatcctgg atgagctgtg
     5941 gcacacaaac gacaatcaga tcgccatctt taaccggctg aagctggtgc caaagaaggt
     6001 ggacctgtcc cagcagaagg agatcccaac cacactggtg gacgatttca tcctgtctcc
     6061 cgtggtgaag cggagcttca tccagagcat caaagtgatc aacgccatca tcaagaagta
     6121 cggcctgccc aatgatatca tcatcgagct ggccagggag aagaactcca aggacgccca
     6181 gaagatgatc aatgagatgc agaagaggaa ccgccagacc aatgagcgga tcgaggagat
     6241 catcagaacc acaggcaagg agaacgccaa gtacctgatc gagaagatca agctgcacga
     6301 tatgcaggag ggcaagtgcc tgtcctacga gacagagatc ctgacagtgg agtatggcct
     6361 gctgccaatc ggcaagatcg tggagaagag gatcgagtgt accgtgtact ctgtggataa
     6421 caatggcaac atctatacac agcccgtggc acagtggcac gataggggag agcaggaggt
     6481 gttcgagtat tgcctggagg acggcagcct gatcagggca accaaggacc acaagttcat
     6541 gacagtggat ggccagatgc tgcccatcga cgagattttc gagcgggagc tggacctgat
     6601 gagagtggat aacctgccta atagcggagg cagtaaaaga acagcagacg ggagtgagtt
     6661 tgagcccaag aaaaagagaa aggtgtaaga tctgataatc aacctctgga ttacaaaatt
     6721 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     6781 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     6841 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6901 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgcgactgtg ccttctagtt
     6961 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     7021 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     7081 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     7141 ggcatgctgg ggatgcggtg ggctctatgg ctcgagaaaa aaatctcgcc aacaagttga
     7201 cgagataaac acggcatttt gccttgtttt agtagattct gtaattttca ttacagagta
     7261 ctaaaaccga gacgcattaa tcgtctcggg tgtttcgtcc tttccacaag atatataaag
     7321 ccaagaaatc gaaatacttt caagttacgg taagcatatg atagtccatt ttaaaacata
     7381 attttaaaac tgcaaactac ccaagaaatt attactttct acgtcacgta ttttgtacta
     7441 atatctttgt gtttacagtc aaattaattc taattatctc tctaacagcc ttgtatcgta
     7501 tatgcaaata tgaaggaatc atgggaaata ggccctcttc ctgcccgacc ttgcggccgc
     7561 aggaacccct agtgatggag ttgg
//
