LOCUS       sequence-401053-        5523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for TRE(TRE3G)-driven smFP_HA (HA-tagged spaghetti 
            monster) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5523)
  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 5523)
  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_HA
FEATURES             Location/Qualifiers
     source          1..5523
                     /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"
     CDS             557..583
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             587..613
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             617..643
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1172..1198
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1205..1231
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1244..1270
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1277..1303
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1514..1540
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1544..1570
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1574..1600
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1650..2238
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1703..1723)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2121..2132)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2270..2746
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2469..2488)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2786..2926
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2786..2915
                     /label=AAV2 ITR
     rep_origin      3001..3456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3088..3107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3298..3319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3473..3492)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3592..3614
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3652..3670)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3738..3842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3843..3911,3912..4703)
                     /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(4061..4080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4874..5462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5363..5382
                     /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 accatgtacc cttatgatgt gcccgattat gctggctacc cttatgatgt
      601 tcctgattac gccggatatc cgtatgacgt gccagactac gcgggaggtg tgagcaaggg
      661 agaggagttg tttaccggcg tggtccccat actggtggag ttggatggcg acgttaatgg
      721 acataaattc tcggtccgcg gcgagggaga gggagacgcc accaatggca agctgacgct
      781 taagttcatt tgcacgactg gaaaattgcc tgtcccctgg ccgaccctgg tcaccacact
      841 gggcggagga gtgcagtgct tctcccgtta cccagaccac atgaagcagc acgatttctt
      901 caagagcgca atgccggagg gatacgtgca agaacggact atttccttca aggatgacgg
      961 aacctataag acccgtgcgg aggtcaaatt cgagggtgac accctggtga accgaattga
     1021 actcaaagga atcgatttca aggaggatgg aaatattctg ggtcacaagc tggaatacaa
     1081 cttcaacagc cataatgtgt acattacggc tgataagcag aagaacggca ttaaggccaa
     1141 tttcaagatc cgccacaacg ttgagggtgg atatccctac gacgtgcccg attatgcggg
     1201 cggatatccg tatgatgttc cagattatgc tggtggtgga ggctatccct atgatgtccc
     1261 cgactacgcc ggaggttacc catacgacgt gcccgactac gctggaggcg atggcagcgt
     1321 gcagctggca gatcattatc aacagaatac ccccataggt gatggccccg ttctgcttcc
     1381 agataatcac tacctttcca cccagagcgt gctttcgaaa gacccgaacg aaaagcgtga
     1441 tcacatggtc ctgctggagt ttgtgaccgc ggctggaatc accctgggta tggacgaact
     1501 ctacaagggt ggctacccct acgatgtgcc ggattacgct ggatatccgt atgacgtacc
     1561 ggactatgcc ggttacccgt acgatgtccc ggactacgct taaaccggtg ctcgaggtac
     1621 ggatccaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     1681 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1741 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1801 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     1861 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     1921 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     1981 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2041 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     2101 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2161 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2221 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     2281 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2341 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     2401 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     2461 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     2521 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     2581 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     2641 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     2701 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     2761 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     2821 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2881 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     2941 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3001 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3061 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3181 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3301 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3361 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3421 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3481 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3541 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3601 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3661 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3721 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3841 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3901 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3961 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4021 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4081 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4141 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4201 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4261 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4321 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4381 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4441 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4501 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4561 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4621 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4681 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4741 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4801 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4861 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4921 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4981 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5041 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5101 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5161 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5221 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5281 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5341 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5401 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5461 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5521 tgt
//
