LOCUS       sequence-394336-        4815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of mTagBFP2 under the TRE promoter in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4815)
  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 4815)
  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-mTagBFP2-WPRE
FEATURES             Location/Qualifiers
     source          1..4815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   51..180
                     /label=AAV2 ITR
     protein_bind    254..272
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    290..308
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    325..343
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    361..379
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    397..415
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    432..450
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    468..486
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        498..536
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     533..557
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      571..580
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             595..1290
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="ELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVVE
                     GGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQ
                     DTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKLV
                     GGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCDL
                     PSKLGHKLN"
     misc_feature    1303..1891
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1356..1376)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1774..1785)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1894..1910)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1895..1911)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    1940..2061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1951..1970)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2005..2024
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2223..2352
                     /label=AAV2 ITR
     rep_origin      complement(2589..3177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2669..2688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3348..4139,4140..4208))
                     /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     3971..3990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4209..4313)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4339..4794)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4476..4497)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4688..4707
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctgcgcgctc
       61 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
      121 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      181 tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct aggaagatct
      241 cgaattggtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
      301 gatagagaac gatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
      361 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta
      421 tgtcgagttt atccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
      481 tagagaacgt atgtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt
      541 tagtgaaccg tcagatcgcc tggagaattc gccaccatgg tgagcaaggg cgaggagctg
      601 attaaggaga acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc
      661 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
      721 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac
      781 ggcagcaaga ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc
      841 cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
      901 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
      961 aacttcacat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc
     1021 gagacgctgt accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc
     1081 gtgggcggga gccatctgat cgcaaacgcc aagaccacat atagatccaa gaaacccgct
     1141 aagaacctca agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag
     1201 gccaacaacg agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc
     1261 cctagcaaac tggggcacaa gctcaattaa aagcttatcg ataatcaacc tctggattac
     1321 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     1381 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     1441 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     1501 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     1561 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     1621 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     1681 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     1741 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     1801 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     1861 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg acccgggcgg
     1921 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1981 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2041 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2101 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     2161 aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     2221 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2281 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     2341 gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2401 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2461 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2521 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2581 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2641 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2701 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2761 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2821 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2881 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2941 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3001 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3061 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3121 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3181 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3241 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3301 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3361 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3421 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3481 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3541 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3601 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3661 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3721 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3781 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3841 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3901 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3961 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4021 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4081 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4141 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4201 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4261 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4321 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4381 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4441 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4501 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4561 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4621 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4681 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4741 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     4801 cgccattcag gctgc
//
