LOCUS       sequence-401052-        5559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for TRE(TRE3G)-driven smFP_Myc (Myc-tagged spaghetti 
            monster) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5559)
  AUTHORS   Watakabe A, Tani T, Abe H, Skibbe H, Ichinohe N, Yamamori T
  TITLE     Serial Two-Photon Tomography of the Whole Marmoset Brain for 
            Neuroanatomical Analyses.
  JOURNAL   J Vis Exp. 2025 Jan 17;(215). doi: 10.3791/67694.
  PUBMED    39895625
REFERENCE   2  (bases 1 to 5559)
  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: "J Vis 
            Exp."; date: "2025-01-17"; volume: "(215). doi"; pages: " 
            10.3791/67694"
COMMENT     Sequence Label: AAVTRE3_smFP_Myc
FEATURES             Location/Qualifiers
     source          1..5559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        147..525
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    155..173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    191..209
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    263..281
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    299..317
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    335..353
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    371..389
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     434..458
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             557..586
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             590..619
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             623..652
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1184..1213
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1220..1249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1262..1291
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1298..1327
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1541..1570
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1574..1603
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1607..1636
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1686..2274
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1739..1759)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2157..2168)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2306..2782
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2505..2524)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2822..2962
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2822..2951
                     /label=AAV2 ITR
     rep_origin      3037..3492
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3124..3143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3334..3355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3509..3528)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3628..3650
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3688..3706)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3774..3878
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3879..3947,3948..4739)
                     /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(4097..4116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4910..5498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5399..5418
                     /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 gcgttcgagt ttactcccta tcagtgatag agaacgtatg
      181 aagagtttac tccctatcag tgatagagaa cgtatgcaga ctttactccc tatcagtgat
      241 agagaacgta taaggagttt actccctatc agtgatagag aacgtatgac cagtttactc
      301 cctatcagtg atagagaacg tatctacagt ttactcccta tcagtgatag agaacgtata
      361 tccagtttac tccctatcag tgatagagaa cgtataagct ttaggcgtgt acggtgggcg
      421 cctataaaag cagagctcgt ttagtgaacc gtcagatcgc ctggagcaat tccacaacac
      481 ttttgtctta taccaacttt ccgtaccact tcctaccctc gtaaagtcga cgaattcaat
      541 tgcggccgcc accatggagc aaaaactgat atcggaggaa gacttggcag agcagaaact
      601 gatttcggaa gaggatcttg ccgaacaaaa actgatcagt gaggaagacc ttgccggagt
      661 gtcgaagggt gaagagctgt tcaccggcgt ggtgcccatc ctcgtcgagc tggacggaga
      721 tgtcaatgga cataaattct ccgtacgggg agagggcgag ggcgacgcaa ccaacggtaa
      781 actgaccctg aagttcattt gcaccacggg taaactgccc gtaccctggc ccaccctggt
      841 cacaaccttg ggtggcggag ttcagtgttt ttcgcggtat ccagatcaca tgaagcagca
      901 tgactttttc aagagcgcta tgcccgaggg ttacgttcaa gagaggacca tcagtttcaa
      961 ggacgacggc acctacaaga cacgggctga ggtgaaattc gagggtgata cactcgtgaa
     1021 tcgtatcgag ctgaagggca tagattttaa ggaggatggc aatatcctgg gccacaagtt
     1081 ggagtataac ttcaacagtc acaatgtcta tataactgcc gataagcaga agaatggcat
     1141 taaagccaat ttcaaaattc gtcacaatgt cgaaggcggc gcagaacaga agctcatcag
     1201 tgaagaggat ctggccgcag agcagaaact cattagcgag gaggacttgg gcggaggcgg
     1261 cgagcagaag ctgatatcgg aggaagacct ggcggccgag caaaagctca tctcggagga
     1321 ggatctcgcg ggaggagatg gaagtgtcca gttggcggat cactaccagc agaatacgcc
     1381 aattggtgac ggccccgtcc ttctgccgga taaccactac ctctccaccc agtcggtgct
     1441 gagcaaggac ccgaatgaaa aacgtgacca catggttctt cttgagttcg tcacggctgc
     1501 gggcatcacg ttgggcatgg acgaactgta caagggcgct gaacagaaac ttatctccga
     1561 ggaggatctg gcggaacaga agctgatcag cgaagaggac ctggcggagc agaaacttat
     1621 cagcgaagag gatctgtaaa ccggtgctcg aggtacggat ccaattcgat atcaagctta
     1681 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1741 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1801 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     1861 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     1921 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     1981 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2041 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     2101 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2161 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2221 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     2281 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     2341 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     2401 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     2461 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     2521 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     2581 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     2641 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     2701 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     2761 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     2821 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2881 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2941 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3001 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3061 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     3121 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3181 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3241 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3301 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3361 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     3421 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3481 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3541 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3601 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3661 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3721 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3781 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3841 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3901 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3961 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4021 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4081 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4141 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4201 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4261 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4321 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4381 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4441 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4501 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4561 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4621 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4681 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4741 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4801 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4981 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5041 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5161 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5221 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5281 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5341 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5401 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5461 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5521 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
