LOCUS       sequence-394375-        4821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of YPet under the TRE promoter in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4821)
  AUTHORS   Leiwe MN, Fujimoto S, Baba T, Moriyasu D, Saha B, Sakaguchi R, 
            Inagaki S, Imai T
  TITLE     Automated neuronal reconstruction with super-multicolour Tetbow 
            labelling and threshold-based clustering of colour hues.
  JOURNAL   Nat Commun. 2024 Jun 25;15(1):5279. doi: 10.1038/s41467-024-49455-y.
  PUBMED    38918382
REFERENCE   2  (bases 1 to 4821)
  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: "Nat 
            Commun."; date: "2024-06-25"; volume: "15(1):5279. doi"; pages: " 
            10.1038/s41467-024-49455-y"
COMMENT     Sequence Label: pAAV-TRE-YPet-WPRE
FEATURES             Location/Qualifiers
     source          1..4821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   65..194
                     /label=AAV2 ITR
     protein_bind    268..286
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    304..322
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    339..357
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    375..393
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    411..429
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    446..464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    482..500
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        512..550
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     547..571
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      585..594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(591..593,594..596,597..1310)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELYK"
     primer_bind     complement(636..657)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(897..916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1323..1911
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1376..1396)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1794..1805)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1914..1930)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1915..1931)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    1960..2081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1971..1990)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2025..2044
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2243..2372
                     /label=AAV2 ITR
     rep_origin      complement(2609..3197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2689..2708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3368..4159,4160..4228))
                     /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     3991..4010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4229..4333)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4359..4814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4496..4517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4708..4727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
       61 ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
      121 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      181 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
      241 ctctaggaag atctcgaatt ggtttactcc ctatcagtga tagagaacgt atgtcgagtt
      301 tactccctat cagtgataga gaacgatgtc gagtttactc cctatcagtg atagagaacg
      361 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
      421 tgatagagaa cgtatgtcga gtttatccct atcagtgata gagaacgtat gtcgagttta
      481 ctccctatca gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg aggcctatat
      541 aagcagagct cgtttagtga accgtcagat cgcctggaga attcgccacc atggtgagca
      601 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      661 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      721 ccctgaaact tctatgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      781 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      841 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      901 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      961 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1021 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1081 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1141 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1201 accagtccgc cctgttcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1261 tcctgaccgc cgccgggatc actgagggca tgaacgagct gtacaagtaa aagcttatcg
     1321 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     1381 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     1441 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     1501 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     1561 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     1621 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1681 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     1741 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     1801 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     1861 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     1921 gtcgacctcg acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     1981 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2041 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2101 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     2161 acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat
     2221 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     2281 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2341 cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg
     2401 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2461 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2521 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2581 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2641 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2701 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2761 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2821 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2881 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2941 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3001 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3061 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3121 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3241 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3301 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3361 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3421 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3481 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3541 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3601 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3661 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3721 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3781 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3841 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3901 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3961 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4021 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4081 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4141 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4201 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4321 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     4381 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4441 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4501 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4561 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4621 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4681 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4741 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4801 gctacagggc gcgtcccatt c
//
