LOCUS       sequence-377169-        6006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector, high-level expression of myrGFP with a 
            somatodendritic-targeting signal (FGL) in neuronal cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6006)
  AUTHORS   Sohn J, Takahashi M, Okamoto S, Ishida Y, Furuta T, Hioki H
  TITLE     A Single Vector Platform for High-Level Gene Transduction of Central
            Neurons: Adeno-Associated Virus Vector Equipped with the Tet-Off 
            System.
  JOURNAL   PLoS One. 2017 Jan 6;12(1):e0169611. doi: 
            10.1371/journal.pone.0169611. eCollection 2017.
  PUBMED    28060929
REFERENCE   2  (bases 1 to 6006)
  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"; journalName: "PLoS One.";
            date: "2017-01-6"; volume: "12(1):e0169611. doi"; pages: " 
            10.1371/journal.pone.0169611. eCollection 2017"
COMMENT     Sequence Label: pAAV2-SynTetOff-FGL
FEATURES             Location/Qualifiers
     source          1..6006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   21..150
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    165..189
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    289..410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(326..345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     380..399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(434..442,443..478,479..481,482..517,
                     518..520,521..556,557..562,563..1180))
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        1701..2015
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1709..1727
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1745..1763
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1780..1798
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1816..1834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1852..1870
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1887..1905
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1923..1941
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1988..2012
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2047..2056
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2110..2119
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2116..2118,2119..2121,2122..2832)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2116..2118,2119..2121,2122..2832)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2161..2182)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2422..2441)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2769..2790
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2995..3219
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(3242..3266)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     repeat_region   3289..3429
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3289..3418
                     /label=AAV2 ITR
     rep_origin      3504..3959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3591..3610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3801..3822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3976..3995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4095..4117
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4155..4173)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4241..4345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4346..4414,4415..5206)
                     /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(4564..4583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5377..5965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5866..5885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
       61 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      121 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtacaagt ttgtacaaaa
      181 aagcaggctt tggatcttac cacatttgta gaggttttac ttgctttaaa aaacctccca
      241 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
      301 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      361 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctga
      421 gatccttact tagttacccg gggagcatgt caaggtcaaa atcgtcaaga gcgtcagcag
      481 gcagcatatc aaggtcaaag tcgtcaaggg catcggctgg gagcatgtct aagtcaaaat
      541 cgtcaagggc gtcggccggc ccgccgcttt cgcactttag ctgtttctcc aggccacata
      601 tgattagttc caggccgaaa aggaaggcag gttcggctcc ctgatggtcg aacagctcaa
      661 ttgcttgtct cagaagtggg ggcatagaat cggtggtagg tgtctctctt tcctcttttg
      721 ctacttgatg ctcctgatcc tccaatacgc agcccagtgt aaagtggccc acggcggaca
      781 gagcgtacag tgcgttctcc agggagaagc cttgctgaca caggaacgcg agctgatttt
      841 ccagggtttc gtactgtttc tctgttgggc gggtgccgag atgcacttta gccccgtcgc
      901 gatgtgagag gagagcacag cggaatgact tggcgttgtt ccgcagaaag tcttgccatg
      961 actcgccttc cagggggcag aagtgggtat gatgcctgtc cagcatctcg attgccaggg
     1021 catcgagcag ggcccgcttg ttcttcacgt gccagtacag ggtaggctgc tcaactccca
     1081 gcttttgagc gagtttcctt gtcgtcaggc cttcgatacc gacttcattg agtaattcca
     1141 gagcagagtt tatgactttg ctcttgtcca gtctagacat ggtgaattcg atatcaagct
     1201 tatcgatacc gtccgccgca gcgcagatgg tcgcgcccgt gcccccctat ctcgcgcctc
     1261 gcgtggtgcg gtccggctgg gccggcggcg gcgcggacgc gaccaaggtg gccgggaagg
     1321 ggagtttgcg ggggaccggc gagtgacgtc agcgcgcctt cagtgctgag gcggcggtgg
     1381 cgcgcgccgc caggcggggg cgaaggcact gtccgcggtg ctgaagctgg cagtgcgcac
     1441 gcgcctcgcc gcatcctgtt tcccctcccc ctctctgata ggggatgcgc aatttgggga
     1501 atgggggttg ggtgcttgtc cagtgggtcg gggtcggtcg tcaggtaggc acccccaccc
     1561 cgcctcatcc tggtcctaaa acccacttgc actcatacgc agggccctct gcagctcgac
     1621 cgcgccgccc cgggcggctc gctgctgccc cctagcgggg gagggacgta attacatccc
     1681 tgggggcttt gggggggctc gagtttactc cctatcagtg atagagaacg tatgtcgagt
     1741 ttactcccta tcagtgatag agaacgatgt cgagtttact ccctatcagt gatagagaac
     1801 gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta ctccctatca
     1861 gtgatagaga acgtatgtcg agtttatccc tatcagtgat agagaacgta tgtcgagttt
     1921 actccctatc agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata
     1981 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag aattcgagct cggtacccgg
     2041 ggatctgcca ccatgggctg tgtgcaatgt aaggataaag aagcaacaaa actgacgggg
     2101 ggatccatcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2161 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     2221 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2281 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     2341 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2401 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2461 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2521 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     2581 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2641 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2701 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     2761 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2821 gagctgtaca agcacaggaa ctggcggctg aagaacatca acagcataaa ctttgacaac
     2881 ccagtctacc agaagaccac agaggacgag ctccacattt gccgaagcca ggatggctat
     2941 acctacccct caagacagat ggtcagcctg gaggacgatg tggcatgaac gcgtctgtgc
     3001 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3061 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3121 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3181 acaatagcag gcatgctggg gatgcggtgg gctctatggg cggccgcgtc gagatatcta
     3241 gacccagctt tcttgtacaa agtggtcacg tgcggaccga gcggccgcag gaacccctag
     3301 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3361 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3421 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3481 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3541 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     3601 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3661 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3721 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3781 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     3841 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3901 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3961 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4021 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4081 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4141 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4201 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4321 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4441 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4501 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4561 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4621 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4741 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4861 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4921 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5521 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6001 cttttg
//
