LOCUS       sequence-268579-        5166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of TRE(TRE3G)-driven, Clover expression for 
            fluorescent labling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5166)
  AUTHORS   Watakabe A, Skibbe H, Nakae K, Abe H, Ichinohe N, Rachmadi MF, Wang 
            J, Takaji M, Mizukami H, Woodward A, Gong R, Hata J, Van Essen DC, 
            Okano H, Ishii S, Yamamori T
  TITLE     Local and long-distance organization of prefrontal cortex circuits 
            in the marmoset brain.
  JOURNAL   Neuron. 2023 May 9:S0896-6273(23)00338-0. doi: 
            10.1016/j.neuron.2023.04.028.
  PUBMED    37196659
REFERENCE   2  (bases 1 to 5166)
  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: "Neuron. 
            2023 May 9:S0896-6273(23)00338-0. doi: 
            10.1016/j.neuron.2023.04.028."
COMMENT     Sequence Label: pAAVTRE3_Clover
FEATURES             Location/Qualifiers
     source          1..5166
                     /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      538..547
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(544..546,547..549,550..1260)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(589..610)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1197..1218
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1293..1881
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1346..1366)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1764..1775)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1913..2389
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2112..2131)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2429..2569
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2429..2558
                     /label=AAV2 ITR
     rep_origin      2644..3099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2731..2750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2941..2962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3116..3135)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3235..3257
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3295..3313)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3381..3485
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3486..3554,3555..4346)
                     /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(3704..3723)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4517..5105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5006..5025
                     /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 cgaattcgcc
      541 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      601 gacggcgacg taaacggcca caagttcagc gtccgcggcg agggcgaggg cgatgccacc
      661 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      721 accctcgtga ccaccttcgg ctacggcgtg gcctgcttca gccgctaccc cgaccacatg
      781 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      841 tctttcaagg acgacggtac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      901 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      961 cacaagctgg agtacaactt caacagccac aacgtctata tcacggccga caagcagaag
     1021 aacggcatca aggctaactt caagatccgc cacaacgttg aggacggcag cgtgcagctc
     1081 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1141 cactacctga gccatcagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1201 gtcctgctgg agttcgtgac cgccgccggg attacacatg gcatggacga gctgtacaag
     1261 taaggatcca attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     1321 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1381 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1441 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     1501 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     1561 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     1621 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     1681 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     1741 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     1801 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     1861 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
     1921 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     1981 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     2041 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     2101 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     2161 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     2221 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     2281 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     2341 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     2401 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     2461 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2521 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     2581 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2641 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2701 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2761 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2821 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2881 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2941 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     3001 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     3061 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     3121 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3181 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3241 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3301 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3361 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3421 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3481 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3541 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3601 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3661 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3721 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3781 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3841 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3901 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3961 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4021 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4081 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4141 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4201 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4261 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4321 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4381 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4441 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4501 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4561 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4621 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4681 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4741 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4801 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4861 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4921 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4981 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5041 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5101 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5161 acatgt
//
