LOCUS       Exported                5871 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  tLuc in AAV backbone.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-tLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5871)
  AUTHORS   Liao HK, Hatanaka F, Araoka T, Reddy P, Wu MZ, Sui Y, Yamauchi T, 
            Sakurai M, O'Keefe DD, Nunez-Delicado E, Guillen P, Campistol JM, Wu
            CJ, Lu LF, Esteban CR, Izpisua Belmonte JC
  TITLE     In Vivo Target Gene Activation via CRISPR/Cas9-Mediated 
            Trans-epigenetic Modulation.
  JOURNAL   Cell. 2017 Dec 14;171(7):1495-1507.e15. doi: 
            10.1016/j.cell.2017.10.025. Epub 2017 Dec 7.
  PUBMED    29224783
REFERENCE   2  (bases 1 to 5871)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5871
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     263..287
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    312..336
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             342..1994
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(408..426)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2128..2249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2165..2184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2219..2238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2476..3064
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2947..2958)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3078..3126
                     /note="synthetic polyadenylation signal"
     repeat_region   3134..3274
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3134..3263
                     /label=AAV2 ITR
     rep_origin      3349..3804
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3436..3455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3646..3667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3821..3840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3940..3962
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4000..4018)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4086..4190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4191..5051
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4409..4428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5222..5810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5711..5730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcctcta gaacgcgtcc tccacggcca ctagatcgat gccctttagg
      181 gataacaggg taatagtgtc ccctccaccc cacagtgcgg gaaactagtt aggcgtgtac
      241 ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagaattc
      301 tgcagatatc aacaagtttg tacaaaaaag caggctccac catggaagac gccaaaaaca
      361 taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga gagcaactgc
      421 ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata
      481 tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg gcagaagcta
      541 tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc
      601 aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg
      661 acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt
      721 tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc
      781 aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt
      841 tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca gagtccttcg
      901 atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt ctgcctaaag
      961 gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga gatcctattt
     1021 ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt
     1081 ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata
     1141 gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa agtgcgctgc
     1201 tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa tacgatttat
     1261 ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc ggggaagcgg
     1321 ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact gagactacat
     1381 cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc
     1441 cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaaa
     1501 gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac aatccggaag
     1561 cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata gcttactggg
     1621 acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag tacaaaggct
     1681 atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac atcttcgacg
     1741 caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt
     1801 tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa
     1861 caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta
     1921 ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa
     1981 agatcgccgt gtaattctag atgtacaagt gaaatcaacc tctggattac aaaatttgtg
     2041 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2101 taatgccttt gtatcatgcg ttaactaaac ttgtttattg cagcttataa tggttacaaa
     2161 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2221 ggtttgtcca aactcatcaa tgtatcttat catgtctgga attgactcaa atgatgtcaa
     2281 ttagtctatc agaagctcat ctggtctccc ttccggggga caagacatcc ctgtttaata
     2341 tttaaacagc agtgttccca aactgggttc ttatatccct tgctctggtc aaccaggttg
     2401 cagggtttcc tgtcctcaca ggaacgaagt ccctaaagaa acagtggcag ccaggtttag
     2461 ccctagtcat atgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2521 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2581 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2641 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2701 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2761 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2821 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     2881 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2941 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3001 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3061 ccgcacaatt ggaattcaat aaaagatctt tattttcatt agatctgtgt gttggttttt
     3121 tgtgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3181 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3241 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3301 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3361 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3421 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3481 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3541 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3601 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3661 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     3721 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3781 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3841 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3901 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3961 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4021 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4081 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4141 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4201 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4261 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4321 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4381 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4441 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4501 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4561 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4621 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4681 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4741 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4801 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4861 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     4921 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4981 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5041 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5101 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5161 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5221 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5281 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5341 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5401 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5461 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5521 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5581 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5641 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5701 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5761 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5821 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
