LOCUS       AAV-CAG-fDIO-ArgiNLS-AausFP1        6240 bp ds-DNA     circular SYN 29-APR-2024
DEFINITION  An AAV genome with Flp recombinase-dependent expression of
            mNeonGreen from the CAG promoter.
ACCESSION
VERSION     .
KEYWORDS    pAAV-CAG-fDIO-mNeonGreen.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct.
REFERENCE   1  (bases 1 to 6156)
  AUTHORS   Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL,
            Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE and Gradinaru
            V
  TITLE     Engineered AAVs for efficient noninvasive gene delivery to the
            central and peripheral nervous systems.
  JOURNAL   Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub
            2017 Jun 26.
  PUBMED    28671695
REFERENCE   2  (bases 1 to 6156)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
COMMENT     ##Imported_From-START##
            Path     :: /Users/ericszelenyi/Downloads
            Filename :: addgene-plasmid-99133-sequence-191157.gbk
            ##Imported_From-END##
FEATURES             Location/Qualifiers
     source          1..6240
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /note="Functional equivalent of wild-type AAV2 ITR"
                     /label="AAV2 ITR (alternate)"
     enhancer        210..513
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        515..792
                     /label="chicken beta-actin promoter"
     primer_bind     1122..1144
                     /note="Chimeric intron in CAG promoter, forward primer"
                     /label="pCAGGS-5"
     primer_bind     1206..1225
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
                     /label="pCAG-F"
     protein_bind    1292..1325
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
                     /label="FRT (minimal)"
     CDS             complement(1418..2137)
                     /created_by="ericszelenyi"
                     /label="AausFP1"
     sig_peptide     complement(2138..2185)
                     /created_by="ericszelenyi"
                     /label="ArgiNLS"
     protein_bind    complement(2225..2258)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
                     /label="FRT (minimal)"
     misc_feature    2367..2955
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /label="WPRE"
     primer_bind     complement(2420..2440)
                     /note="WPRE, reverse primer"
                     /label="WPRE-R"
     CDS             complement(2838..2849)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
                     /label="Factor Xa site"
     polyA_signal    2987..3463
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     primer_bind     complement(3186..3205)
                     /note="Human growth hormone terminator, reverse primer"
                     /label="hGH-PA-R"
     repeat_region   3503..3643
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /label="AAV2 ITR"
     repeat_region   3503..3632
                     /label="AAV2 ITR"
     rep_origin      3718..4173
                     /direction="RIGHT"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     primer_bind     complement(3805..3824)
                     /note="F1 origin, reverse primer"
                     /label="F1ori-R"
     primer_bind     4015..4036
                     /note="F1 origin, forward primer"
                     /label="F1ori-F"
     primer_bind     complement(4190..4209)
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /label="pRS-marker"
     primer_bind     4309..4331
                     /note="pGEX vectors, reverse primer"
                     /label="pGEX 3'"
     primer_bind     complement(4369..4387)
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
                     /label="pBRforEco"
     promoter        4455..4559
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4560..5420
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="AmpR"
     primer_bind     complement(4778..4797)
                     /note="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     rep_origin      5591..6179
                     /direction="RIGHT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     6080..6099
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
ORIGIN      
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaacact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg
      661 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga
     1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa
     1261 cggtaccgga tcctctagag tcgactccgg agaagttcct attctctaga aagtatagga
     1321 acttcgcaga atggtagctg gattgtagct gctattagca atatgaaacc tcttagaagt
     1381 tcctattctt caaaaggtat aggaacttcg gcgcgcctta ggcgtaggcg gtcttgcagt
     1441 cgatggctct gaacacttcc accacgttca tgtggtcccg ggtttcttta gggtccttgc
     1501 tcaggatggt gtgggcctga atgtggtgat agtgagggat gtccacgggg ccacctccga
     1561 gaggggtatt cagctgctgg tgagcagcaa tgatctggcc tccgccgatc acgtcgtgca
     1621 caatgttgat gtagcacttg atgccgttgt tctgcacgtc gggcagcacg tagatgcagt
     1681 gcgggttgta gttgaactca agtttcttgc ccaggatgtt gccgcctttc ttgaagccgc
     1741 tgccgttcag ggtcactctg ttgtacacgg agcccttctc gtaggtcact tcggctctgg
     1801 tcttgtacac gccgtcgttc tcgaaggtga tggttctctc ttgcacgtag ccctcaggca
     1861 tggcgctctt ggggtagtcc ttgatgtcgt tgggatactt ggcgaagcac tgggcgccgt
     1921 atgtcactgt ggtcaggatg ctgctccaag gcacaggcag ttcgccggtt gtgcagatga
     1981 agctggcctc gatctttccg gtattggcgt cgccgtgtcc tttgcctctg acgctgaact
     2041 tcatgccttc cacgtcgcct tccatttcca ccacataggg gatcttctcg cggaacagca
     2101 gggcgccata agacatggtg gccacagggg atcccgttcg acggcgccgc cgccttctca
     2161 ccttcttttt ctttgctatg ctcatctgct tctggggagc cacccttttc atggtggcgc
     2221 tagcgaagtt cctatacttt ctagagaata ggaacttctt gccttaaccc agaaattatc
     2281 actgttattc tttagaatgg tgcaaagaga agttcctata ccttttgaag aataggaact
     2341 tcgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2401 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2461 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2521 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2581 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2641 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2701 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2761 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     2821 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2881 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2941 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3001 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3061 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3121 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3181 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3241 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3301 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3361 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3421 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     3481 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3541 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3601 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3661 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3721 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3781 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3841 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3901 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3961 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4021 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4081 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4141 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4201 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4261 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4321 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4381 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4441 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4501 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4561 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4621 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4681 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4741 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4801 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4861 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4921 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4981 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5041 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5101 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5161 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5221 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5281 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5341 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5401 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5461 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5521 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5581 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5641 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5701 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5761 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5821 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5881 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5941 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6001 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6061 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6121 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6181 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
