LOCUS       sequence-191158-        5882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome that ubiquitously expresses the fluorescent protein 
            mNeonGreen from the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5882)
  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 5882)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nn.4593"; 
            journalName: "Nat Neurosci"; date: "2017-08"; volume: "20"; issue: 
            "8"; pages: "1172-1179"
COMMENT     Sequence Label: pAAV-CAG-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..5882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1274..1984
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1529..1538
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2009..2597
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2062..2082)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2480..2491)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2629..3105
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2828..2847)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3145..3285
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3145..3274
                     /label=AAV2 ITR
     rep_origin      3360..3815
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3447..3466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3657..3678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3832..3851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3951..3973
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4011..4029)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4097..4201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4202..4270,4271..5062)
                     /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(4420..4439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5233..5821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5722..5741
                     /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 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 cggtaccgcc accatggtga gcaagggcga ggaggataac atggcctctc tcccagcgac
     1321 acatgagtta cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg
     1381 caccggcaat ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct
     1441 ccagttctcc ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc
     1501 ctaccctgac gggatgtcgc ctttccaggc cgccatggta gatggctccg gataccaagt
     1561 ccatcgcaca atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta
     1621 cgagggaagc cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg
     1681 tcctgtgatg accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc
     1741 caacgacaaa accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg
     1801 ctaccggagc actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct
     1861 gaagaaccag ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct
     1921 caacttcaag gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa
     1981 gtgagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     2041 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2101 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2161 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2221 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2281 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2341 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2401 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     2461 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2521 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2581 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc
     2641 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     2701 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     2761 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     2821 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     2881 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     2941 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     3001 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     3061 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     3121 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3181 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3241 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3301 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3361 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3421 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3481 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3541 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     3601 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3661 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     3721 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3781 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3841 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3901 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3961 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4021 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4081 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4141 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4201 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4261 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4321 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4381 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4441 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4501 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4561 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4621 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4681 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4741 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4801 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4861 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4921 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4981 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5041 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5101 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5161 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5221 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5281 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5341 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5401 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5461 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5521 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5581 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5641 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5701 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5761 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5821 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5881 gt
//
