LOCUS       sequence-401056-        5501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of TRE(TRE3G)-driven, Vamp2-conjugated mTFP1 
            expression for fluorescent labling of axon terminals.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5501)
  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 5501)
  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: pAAV-TRE3_ mTFP1- Vamp2
FEATURES             Location/Qualifiers
     source          1..5501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     144..161
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     145..161
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        162..540
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    170..188
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    242..260
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    278..296
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    314..332
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    350..368
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    386..404
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     449..473
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      550..559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             556..1263
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     misc_feature    1655..2243
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1708..1728)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2126..2137)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2249..2725
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2448..2467)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2764..2904
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2764..2893
                     /label=AAV2 ITR
     rep_origin      2979..3434
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3066..3085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3276..3297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3451..3470)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3570..3592
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3630..3648)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3716..3820
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3821..3889,3890..4681)
                     /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(4039..4058)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4852..5440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5341..5360
                     /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 gcggcccgtc gactgtaaaa cgacggccag tgagtttact ccctatcagt
      181 gatagagaac gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta
      241 ctccctatca gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt
      301 atgaccagtt tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt
      361 gatagagaac gtatatccag tttactccct atcagtgata gagaacgtat aagctttagg
      421 cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt gaaccgtcag atcgcctgga
      481 gcaattccac aacacttttg tcttatacca actttccgta ccacttccta ccctcgtaaa
      541 gctagagccg ccaccatggt gagcaagggc gaggagacca caatgggcgt aatcaagccc
      601 gacatgaaga tcaagctgaa gatggagggc aacgtgaatg gccacgcctt cgtgatcgag
      661 ggcgagggcg agggcaagcc ctacgacggc accaacacca tcaacctgga ggtgaaggag
      721 ggagcccccc tgcccttctc ctacgacatt ctgaccaccg cgttcgccta cggcaacagg
      781 gccttcacca agtaccccga cgacatcccc aactacttca agcagtcctt ccccgagggc
      841 tactcttggg agcgcaccat gaccttcgag gacaagggca tcgtgaaggt gaagtccgac
      901 atctccatgg aggaggactc cttcatctac gagatacacc tcaagggcga gaacttcccc
      961 cccaacggcc ccgtgatgca gaagaagacc accggctggg acgcctccac cgagaggatg
     1021 tacgtgcgcg acggcgtgct gaagggcgac gtcaagcaca agctgctgct ggagggcggc
     1081 ggccaccacc gcgttgactt caagaccatc tacagggcca agaaggcggt gaagctgccc
     1141 gactatcact ttgtggacca ccgcatcgag atcctgaacc acgacaagga ctacaacaag
     1201 gtgaccgttt acgagagcgc cgtggcccgc aactccaccg acggcatgga cgagctgtac
     1261 aaggctagct cggctaccgc tgccaccgtc ccgcctgccg ccccggccgg cgagggtggc
     1321 ccccctgcac ctcctccaaa ccttactagt aacaggagac tgcagcagac ccaggcccag
     1381 gtggatgagg tggtggacat catgagggtg aatgtggaca aggtcctgga gcgggaccag
     1441 aagttgtcgg agctggatga ccgtgcagat gccctccagg caggggcctc ccagtttgaa
     1501 acaagtgcag ccaagctcaa gcgcaaatac tggtggaaaa acctcaagat gatgatcatc
     1561 ttgggagtga tctgcgccat catcctcatc atcatcatcg tttacttcag cacttaagcg
     1621 gccgcggatc agtcgaggga tcgagcttat cgataatcaa cctctggatt acaaaatttg
     1681 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1741 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1801 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1861 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1921 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     1981 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2041 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     2101 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2161 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2221 ctccctttgg gccgcctccc cgcatcacgg gtggcatccc tgtgacccct ccccagtgcc
     2281 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     2341 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     2401 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     2461 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     2521 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     2581 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     2641 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     2701 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagggc
     2761 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2821 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2881 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2941 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3001 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     3061 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3121 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3181 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3241 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3301 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3361 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3421 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3481 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3541 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3601 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3661 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3721 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3781 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3841 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3901 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3961 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4021 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4081 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4141 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4201 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4261 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4321 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4381 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4441 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4501 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4561 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4621 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4681 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4741 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4801 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4861 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4921 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4981 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5041 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5101 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5161 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5221 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5281 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5341 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5401 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5461 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
