LOCUS       Exported                7131 bp ds-DNA     circular SYN 31-JAN-2023
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 7131)
  AUTHORS   Ye L, Wang J, Tan Y, Beyer AI, Xie F, Muench MO, 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 7131)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Feb 1, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7131
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3879..4529
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    122..282
                     /standard_name="lacZ_a reporter"
                     /label=lacZ_a reporter
                     /note="Geneious type: reporter"
     misc_feature    300..606
                     /standard_name="f1 origin"
                     /label=f1 origin
                     /note="Geneious type: origin of replication"
     misc_feature    729
                     /standard_name="T"
                     /label=T
                     /note="Geneious type: Editing History Replacement"
     misc_feature    733
                     /standard_name="T"
                     /label=T
                     /note="Geneious type: Editing History Replacement"
     misc_feature    738^739
                     /standard_name="C"
                     /label=C
                     /note="Geneious type: Editing History Deletion"
     misc_feature    747..748
                     /standard_name="TC"
                     /label=TC
                     /note="Geneious type: Editing History Replacement"
     regulatory      799..827
                     /standard_name="amp prom"
                     /label=amp prom
                     /note="Geneious type: promoter"
     misc_feature    869..1729
                     /standard_name="amp marker"
                     /label=amp marker
                     /note="Geneious type: marker"
     CDS             869..1729
                     /standard_name="Amp-R"
                     /label=Amp-R
     misc_feature    1416
                     /standard_name="T"
                     /label=T
                     /note="Geneious type: Editing History Replacement"
     misc_feature    1422
                     /standard_name="T"
                     /label=T
                     /note="Geneious type: Editing History Replacement"
     misc_feature    1884..2503
                     /standard_name="pBR322 origin"
                     /label=pBR322 origin
                     /note="Geneious type: origin of replication"
     misc_feature    2047
                     /standard_name="C"
                     /label=C
                     /note="Geneious type: Editing History Replacement"
     misc_feature    2533..2535
                     /standard_name="CGG"
                     /label=CGG
                     /note="Geneious type: Editing History Replacement"
     regulatory      2812..2841
                     /standard_name="lac prom"
                     /label=lac prom
                     /note="Geneious type: promoter"
     repeat_region   2917..3046
                     /standard_name="5'-ITR pSub201(+)"
                     /label=5'-ITR pSub201(+)
     regulatory      3063..3866
                     /standard_name="CBh promoter"
                     /label=CBh promoter
                     /note="Geneious type: promoter"
     regulatory      3071..3356
                     /standard_name="CMV Enhancer"
                     /label=CMV Enhancer
                     /note="Geneious type: promoter"
     regulatory      3369..3630
                     /standard_name="Chicken B-actin promoter"
                     /label=Chicken B-actin promoter
                     /note="Geneious type: promoter"
     regulatory      3607..3612
                     /standard_name="CBA TATA Box"
                     /label=CBA TATA Box
                     /note="Geneious type: promoter"
     regulatory      3631..3769
                     /standard_name="Chicken B-actin intron"
                     /label=Chicken B-actin intron
                     /note="Geneious type: promoter"
     regulatory      3637
                     /standard_name="putative cba +1"
                     /label=putative cba +1
                     /note="Geneious type: promoter"
     misc_feature    3645^3646
                     /standard_name="A"
                     /label=A
                     /note="Geneious type: Editing History Deletion"
     regulatory      3724..3732
                     /standard_name="b-actin SD (MaxEnt)"
                     /label=b-actin SD (MaxEnt)
                     /note="Geneious type: promoter"
     regulatory      3775..3831
                     /standard_name="MVM Intron"
                     /label=MVM Intron
                     /note="Geneious type: promoter"
     regulatory      3777..3785
                     /standard_name="MVM SD"
                     /label=MVM SD
                     /note="Geneious type: promoter"
     regulatory      3832..3866
                     /standard_name="MVM Intron 2"
                     /label=MVM Intron 2
                     /note="Geneious type: promoter"
     regulatory      complement(3859..3863)
                     /standard_name="MVM SA (MaxEnt)"
                     /label=MVM SA (MaxEnt)
                     /note="Geneious type: promoter"
     misc_feature    3873..3882
                     /standard_name="Kozak"
                     /label=Kozak
                     /note="Geneious type: motif"
     CDS             3879..3935
                     /standard_name="BPNLS"
                     /label=BPNLS
     CDS             3933..4433
                     /standard_name="TadA*8e"
                     /label=TadA*8e
     misc_feature    3999..4001
                     /standard_name="W23R"
                     /label=W23R
     misc_feature    4038..4040
                     /standard_name="H36L"
                     /label=H36L
     misc_feature    4074..4076
                     /standard_name="P48A"
                     /label=P48A
     misc_feature    4083..4085
                     /standard_name="R51L"
                     /label=R51L
     misc_feature    4182..4184
                     /standard_name="L84F"
                     /label=L84F
     misc_feature    4248..4250
                     /standard_name="A106V"
                     /label=A106V
     misc_feature    4254..4256
                     /standard_name="D108N"
                     /label=D108N
     misc_feature    4257..4259
                     /standard_name="A109S"
                     /label=A109S
     misc_feature    4263..4265
                     /standard_name="T111R"
                     /label=T111R
     misc_feature    4287..4289
                     /standard_name="D119N"
                     /label=D119N
     misc_feature    4296..4298
                     /standard_name="H122N"
                     /label=H122N
     misc_feature    4299..4301
                     /standard_name="H123Y"
                     /label=H123Y
     misc_feature    4368..4370
                     /standard_name="S146C"
                     /label=S146C
     misc_feature    4371..4373
                     /standard_name="D147 (changed back from D147Y in ABE7.10)"
                     /label=D147 (changed back from D147Y in ABE7.10)
     misc_feature    4377..4379
                     /standard_name="F149Y"
                     /label=F149Y
     misc_feature    4386..4388
                     /standard_name="R152P"
                     /label=R152P
     misc_feature    4395..4397
                     /standard_name="E155V"
                     /label=E155V
     misc_feature    4398..4400
                     /standard_name="I156F"
                     /label=I156F
     misc_feature    4401..4403
                     /standard_name="K157N"
                     /label=K157N
     misc_feature    4428..4430
                     /standard_name="T166I"
                     /label=T166I
     misc_feature    4431..4433
                     /standard_name="D167N"
                     /label=D167N
     CDS             4434..4529
                     /standard_name="linker"
                     /label=linker
     CDS             4527..6245
                     /standard_name="nSpCas9(D10A/A61R)"
                     /label=nSpCas9(D10A/A61R)
                     /note="hSpCas9"
     misc_difference 4554..4556
                     /standard_name="D10A"
                     /label=D10A
     misc_difference 4707..4709
                     /standard_name="A61R (GCC>AGA)"
                     /label=A61R (GCC>AGA)
     CDS             6246..6551
                     /standard_name="NpuN"
                     /label=NpuN
     CDS             6552..6563
                     /standard_name="linker"
                     /label=linker
     CDS             6564..6614
                     /standard_name="BPNLS"
                     /label=BPNLS
     CDS             6615..6617
     regulatory      6623..6783
                     /standard_name="WPRE_Gamma"
                     /label=WPRE_Gamma
                     /note="Geneious type: terminator"
     regulatory      6872..7099
                     /standard_name="bGH_PA term"
                     /label=bGH_PA term
                     /note="Geneious type: Terminator"
     repeat_region   join(7108..7131,1..106)
                     /standard_name="3'-ITR pSub201(+)"
                     /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 ctgcgcgctg ccttcgcccc
     3661 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     3721 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
     3781 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
     3841 cctgaaatca ctttttttca ggttggaccg gtgccaccat gaaacggaca gccgacggaa
     3901 gcgagttcga gtcaccaaag aagaagcgga aagtctctga ggtggagttt tcccacgagt
     3961 actggatgag acatgccctg accctggcca agagggcacg cgatgagagg gaggtgcctg
     4021 tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac agagccatcg
     4081 gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc ggcctggtca
     4141 tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct tgcgtgatgt
     4201 gcgccggcgc catgatccac tctaggatcg gccgcgtggt gtttggcgtg aggaacagca
     4261 aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg aatcaccgcg
     4321 tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc gatttctacc
     4381 ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc aactccggag
     4441 gatctagcgg aggctcctct ggctctgaga cacctggcac aagcgagagc gcaacacctg
     4501 aaagcagcgg gggcagcagc ggggggtcag acaagaagta cagcatcggc ctggccatcg
     4561 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     4621 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     4681 tgttcgacag cggcgaaaca gccgagagaa cccggctgaa gagaaccgcc agaagaagat
     4741 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     4801 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     4861 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     4921 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     4981 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     5041 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     5101 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     5161 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     5221 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     5281 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     5341 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     5401 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     5461 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     5521 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     5581 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     5641 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     5701 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     5761 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     5821 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     5881 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     5941 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     6001 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     6061 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     6121 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     6181 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     6241 tcgagtgcct gtcctacgag acagagatcc tgacagtgga gtatggcctg ctgccaatcg
     6301 gcaagatcgt ggagaagagg atcgagtgta ccgtgtactc tgtggataac aatggcaaca
     6361 tctatacaca gcccgtggca cagtggcacg ataggggaga gcaggaggtg ttcgagtatt
     6421 gcctggagga cggcagcctg atcagggcaa ccaaggacca caagttcatg acagtggatg
     6481 gccagatgct gcccatcgac gagattttcg agcgggagct ggacctgatg agagtggata
     6541 acctgcctaa tagcggaggc agtaaaagaa cagcagacgg gagtgagttt gagcccaaga
     6601 aaaagagaaa ggtgtaagat ctgataatca acctctggat tacaaaattt gtgaaagatt
     6661 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     6721 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     6781 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     6841 tcggctgttg ggcactgaca attccgtggt gcgactgtgc cttctagttg ccagccatct
     6901 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6961 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     7021 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     7081 gatgcggtgg gctctatggg cggccgcagg aacccctagt gatggagttg g
//
