LOCUS       Exported                5885 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  An AAV genome that ubiquitously expresses the fluorescent protein 
            mRuby2 from the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CAG-mRuby2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5885)
  AUTHORS   Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, 
            Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, 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 5885)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5885
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        210..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..792
                     /label=chicken beta-actin promoter
     primer_bind     1122..1144
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1206..1225
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1268..1277
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1274..1984
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    2012..2600
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2065..2085)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2483..2494)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2632..3108
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2831..2850)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3148..3288
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3148..3277
                     /label=AAV2 ITR
     rep_origin      3363..3818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3450..3469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3660..3681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3835..3854)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3954..3976
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4014..4032)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4100..4204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4205..5065
                     /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(4423..4442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5236..5824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5725..5744
                     /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 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 tgcgcgcggc 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 cggtaccgcc accatggtga gcaagggaga ggaactcatc aaggagaaca tgcgcatgaa
     1321 ggtagtcatg gaagggtctg tgaatgggca tcagttcaag tgcacagggg aaggtgaagg
     1381 gaatccctac atgggtactc agaccatgag gatcaaggtt attgaaggtg gccctctccc
     1441 gtttgccttc gacattcttg ccactagctt catgtatggc agtcggacct tcatcaagta
     1501 ccccaaaggc attccagact tcttcaaaca gtcctttcca gagggcttca catgggaaag
     1561 ggtgacccgt tacgaggatg gtggtgtggt cacagtcatg caggacacta gtctggagga
     1621 cggatgtctg gtctatcacg tgcaagtgag aggggtgaac tttccctcca atgggcctgt
     1681 gatgcagaag aaaaccaaag ggtgggagcc aaacacagag atgatgtatc ctgcagatgg
     1741 aggactgaga ggctatacgc acatggctct gaaggttgac ggaggaggcc acttgtcatg
     1801 ctcatttgtg accacgtaca ggagcaagaa aactgtcggc aacatcaaaa tgcctggcat
     1861 acatgcggtt gatcaccgct tggaacgact ggaggagtct gacaacgaga tgtttgtggt
     1921 acaacgggaa catgcagttg ccaagtttgc tggccttggc ggaggaatgg atgagctgta
     1981 caaatgagaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     2041 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2101 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2161 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2221 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2281 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2341 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2401 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     2461 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2521 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2581 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     2641 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     2701 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     2761 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     2821 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     2881 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     2941 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     3001 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     3061 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     3121 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3181 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3241 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3301 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3361 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3421 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3481 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3541 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     3601 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3661 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3721 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3781 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3841 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3901 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3961 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4021 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4081 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4141 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4201 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4261 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4321 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4381 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4441 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4501 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4561 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4621 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4681 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4741 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4801 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4861 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4921 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4981 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5041 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5101 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5161 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5221 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5281 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5341 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5401 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5461 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5521 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5581 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5641 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5701 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5761 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5821 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5881 catgt
//
