LOCUS       Exported                4596 bp ds-DNA     circular SYN 15-FEB-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    1183_pAAV-U6-Ex14-gRNAd-CB-EmGFP_89058
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4596)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Feb 15, 2017 from SnapGene 2.7.3
            http://www.snapgene.com
COMMENT     Component Fragments
            #1: FRAGMENT of pZAC2.1rhEPO.36.Q-L
             parent position: from 1309 to 239
             original length: 3894
             molecule position: from 1 to 3894
            Left Terminus
              XhoI site #1
            Right Terminus
              BglII site #1
            #2: FRAGMENT of pAAVNEWTBG-BI-aGAL
             parent position: from 5114 to 316
             original length: 896
             molecule position: from 3895 to 4790
            Left Terminus
              BamHI site #1
            Right Terminus
              XhoI site #1
            http://www.informaxinc.com/
FEATURES             Location/Qualifiers
     source          1..4596
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..185
                     /label=ITR
                     /note="5'ITR"
     promoter        255..495
                     /label=U6 Promoter
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     276..310
                     /note="U6ForSeq"
     misc_feature    504..523
                     /note="Ex14gRNA-d"
     misc_feature    524..600
                     /note="guideRNA backbone"
     misc_feature    622..627
                     /note="Cloning Site for Additional Guide RNA cassettes"
     misc_feature    628..1086
                     /note="Promoter cassette from pAAV-CB-EGFP"
     enhancer        683..808
                     /note="Partial CMV enhancer"
     misc_feature    complement(696..717)
                     /note="CBP Rev3"
     promoter        809..1039
                     /note="CB promoter"
     TATA_signal     1015..1022
                     /note="TATA"
     CDS             1090..1809
                     /codon_start=1
                     /product="Emerald GFP"
                     /note="EmGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1855..1915
                     /note="Syn PolyA"
     primer_bind     complement(1879..1908)
                     /note="SynPolyARevSeq"
     misc_feature    1916..2088
                     /label=ITR
                     /note="3'ITR"
     misc_feature    complement(2113..4528)
                     /label=pEMBL8 Plasmid Backbone
                     /note="pEMBL8 Plasmid Backbone"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctggtgtac aaaaaagcag gctttaaagg aaccaattca gtcgactgga tccggtacca
      241 aggtcgggca ggaagagggc ctatttccca tgattccttc atatttgcat atacgataca
      301 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
      361 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
      421 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
      481 tcttgtggaa aggacgaaac acctgctgct ggccaaggac atggttttag agctagaaat
      541 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      601 ttttttctag acccagcttt caagcttgat taacccgcca tgctacttat ctacgtagcc
      661 atgctctagg aagatcggaa ttcgccctta agctagtatg ccaagtacgc cccctattga
      721 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      781 tcctacttgg cagtacatct actcgaggcc acgttctgct tcactctccc catctccccc
      841 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      901 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
      961 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
     1021 aagcgaagcg cgcggcgggc gggagcggga tcagaatgat ctgatatcat cgatgaattc
     1081 gagctcacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1141 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1201 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1261 tggcccaccc tcgtgaccac cttcacctac ggcgtgcagt gcttcgcccg ctaccccgac
     1321 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1381 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1441 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1501 ctggggcaca agctggagta caactacaac agccacaagg tctatatcac cgccgacaag
     1561 cagaagaacg gcatcaaggt gaacttcaag acccgccaca acatcgagga cggcagcgtg
     1621 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1681 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1741 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1801 tacaagtaag aattctgcag tcgacgcggg cccgggatcc accggtgcgg ccgctcgcga
     1861 ataaaagatc tttattttca ttagatctgt gtgttggttt tttgtgtgat gcagctacgt
     1921 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     1981 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2041 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     2101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaccgcag
     2281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2761 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3361 ccagttaata gtttgcgcaa cgttgttacc attactacag gcatcgtggt gtcacgctcg
     3421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3781 atcttaccac tattgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa agatgccacc tgaaattata
     4081 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4141 caataggccg aaatcggaaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4201 agtgttgttc cagtttggaa caagagtcca ctattgagga acgtgaactc cagcgtcaaa
     4261 gggcgaaaaa ccgtctatcg gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4321 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4381 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4441 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4501 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     4561 gaagggcgat cggtgcgggc ctcttcgcta ttacgc
//
