LOCUS       sequence-190885-        5451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome with tet-inducible, Cre-dependent expression of the 
            fluorescent protein mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5451)
  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 5451)
  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-TRE-DIO-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..5451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    165..183
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    202..220
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    239..257
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    276..294
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    313..331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    350..368
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    387..405
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    424..442
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     464..488
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(587..620)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(671..704)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(713..1423)
                     /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      1159..1168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1420..1429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1436..1469
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1520..1553
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1578..2166
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1631..1651)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2049..2060)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2198..2674
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2397..2416)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2714..2854
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2714..2843
                     /label=AAV2 ITR
     rep_origin      2929..3384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3016..3035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3226..3247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3401..3420)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3520..3542
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3580..3598)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3666..3770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3771..3839,3840..4631)
                     /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(3989..4008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4802..5390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5291..5310
                     /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 gcgtatcaag tgccacctga cgtctcccta tcagtgatag
      181 agaagtcgac acgtctcgag ctccctatca gtgatagaga aggtacgtct agaacgtctc
      241 cctatcagtg atagagaagt cgacacgtct cgagctccct atcagtgata gagaaggtac
      301 gtctagaacg tctccctatc agtgatagag aagtcgacac gtctcgagct ccctatcagt
      361 gatagagaag gtacgtctag aacgtctccc tatcagtgat agagaagtcg acacgtctcg
      421 agctccctat cagtgataga gaagctaccc cctatataag cagagctcgt ttagtgaacc
      481 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      541 gatccagcct ggatccggta ccggatcctc tagagtcgac tccggaataa cttcgtatag
      601 gatactttat acgaagttat gcagaatggt agctggattg tagctgctat tagcaatatg
      661 aaacctctta ataacttcgt atagcataca ttatacgaag ttatggcgcg cctcacttgt
      721 acagctcgtc catgcccatc acatcggtaa aggccttttg ccactccttg aagttgagct
      781 cggtcttgga gtgcttgagc tccgtcttac ggaacacgta catcggctgg ttcttcagat
      841 agttagccgc cattggcttg gcaaaggtgt aggtggtccg cgcagtgctc cggtagcgct
      901 tgccatttcc agtggtgtaa ctccacttaa aggtactgat gatggttttg tcgttggggt
      961 aagtcttctt cgacctgcac cagtccgcag cggtcagcga gttggtcatc acaggaccgt
     1021 cagcagggaa accagtcccc ttcacctggg cctctccttt gatgtggctt ccctcgtagg
     1081 tgtagcggta gttaacagta agggaggcac catcttcaaa ctgcattgtg cgatggactt
     1141 ggtatccgga gccatctacc atggcggcct ggaaaggcga catcccgtca gggtagggca
     1201 ggtactgatg gaagccatac ccgatatgag ggaccagaat ccagggggag aactggaggt
     1261 cacccttggt ggacttcagg tttaactcct cataaccatc atttggattg ccggtgccct
     1321 gacccaccat gtcaaagtcc acaccgttga tggagccaaa gatgtgtaac tcatgtgtcg
     1381 ctgggagaga ggccatgtta tcctcctcgc ccttgctcac catggtggcg ctagcataac
     1441 ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt
     1501 ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg
     1561 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1621 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1681 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     1741 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     1801 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     1861 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1921 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     1981 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     2041 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2101 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2161 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     2221 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     2281 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     2341 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     2401 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     2461 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     2521 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     2581 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     2641 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     2701 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2761 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2821 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2881 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2941 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3001 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3061 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3121 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3181 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3241 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     3301 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3361 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3421 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3481 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3541 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3601 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3661 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3721 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3781 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3841 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3901 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3961 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4021 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4081 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4141 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4201 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4261 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4321 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4381 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4441 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4501 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4561 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4621 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4681 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4741 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4801 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4861 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4921 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4981 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5041 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5101 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5161 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5221 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5281 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5341 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5401 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
