LOCUS       Exported                5524 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  An AAV genome encoding expression of the nuclear localized 
            fluorescent protein mTurquoise2 from the GFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-GFAP-2xNLS-mTurquoise2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5524)
  AUTHORS   Challis RC, Ravindra Kumar S, Chan KY, Challis C, Beadle K, Jang MJ,
            Kim HM, Rajendran PS, Tompkins JD, Shivkumar K, Deverman BE, 
            Gradinaru V
  TITLE     Systemic AAV vectors for widespread and targeted gene delivery in 
            rodents.
  JOURNAL   Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3.
  PUBMED    30626963
REFERENCE   2  (bases 1 to 5524)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5524
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      832..841
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             847..867
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      874..883
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             880..1596
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             880..1596
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(925..946)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1186..1205)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1533..1554
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1603..1623
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1651..2239
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1704..1724)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2122..2133)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2271..2747
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2470..2489)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2787..2927
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2787..2916
                     /label=AAV2 ITR
     rep_origin      3002..3457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3089..3108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3299..3320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3474..3493)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3593..3615
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3653..3671)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3739..3843
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3844..4704
                     /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(4062..4081)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4875..5463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5364..5383
                     /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 gcgttaacat atcctggtgt ggagtagggg acgctgctct
      181 gacagaggct cgggggcctg agctggctct gtgagctggg gaggaggcag acagccaggc
      241 cttgtctgca agcagacctg gcagcattgg gctggccgcc ccccagggcc tcctcttcat
      301 gcccagtgaa tgactcacct tggcacagac acaatgttcg gggtgggcac agtgcctgct
      361 tcccgccgca ccccagcccc cctcaaatgc cttccgagaa gcccattgag cagggggctt
      421 gcattgcacc ccagcctgac agcctggcat cttgggataa aagcagcaca gccccctagg
      481 ggctgccctt gctgtgtggc gccaccggcg gtggagaaca aggctctatt cagcctgtgc
      541 ccaggaaagg ggatcagggg atgcccaggc atggacagtg ggtggcaggg ggggagagga
      601 gggctgtctg cttcccagaa gtccaaggac acaaatgggt gaggggagag ctctccccat
      661 agctgggctg cggcccaacc ccaccccctc aggctatgcc agggggtgtt gccaggggca
      721 cccgggcatc gccagtctag cccactcctt cataaagccc tcgtatccca ggagcgagca
      781 gagccagagc aggttggaga ggagacgcat cacctctgct gcttgggtac cgccaccatg
      841 ggaagcccaa agaagaagcg taaggtagga tccgccacca tggtgagcaa gggcgaggag
      901 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      961 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1021 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgtcctgg
     1081 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1141 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1201 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1261 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacttc
     1321 agcgacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1381 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1441 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     1501 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1561 gccgggatca ctctcggcat ggacgagctg tacaagggaa gccccaagaa aaagcggaag
     1621 gtgtgagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     1681 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     1741 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     1801 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     1861 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     1921 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     1981 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2041 gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg
     2101 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2161 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2221 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc
     2281 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     2341 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     2401 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     2461 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     2521 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     2581 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     2641 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     2701 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg
     2761 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     2821 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     2881 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     2941 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     3001 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3061 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3181 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     3241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3301 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     3361 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     3421 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     3481 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     3541 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3601 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     3661 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3721 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3781 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3841 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3901 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3961 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4021 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4081 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4141 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4201 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4261 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4321 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4381 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4441 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4501 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4561 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4621 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4681 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4741 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4801 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4861 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4921 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4981 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5041 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5101 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5161 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5221 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5281 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5341 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5401 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5461 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5521 atgt
//
