LOCUS       sequence-227884-        5638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for sparse and bright labeling in a cell-type specific 
            manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5638)
  AUTHORS   Lin R, Wang R, Yuan J, Feng Q, Zhou Y, Zeng S, Ren M, Jiang S, Ni H,
            Zhou C, Gong H, Luo M
  TITLE     Cell-type-specific and projection-specific brain-wide reconstruction
            of single neurons.
  JOURNAL   Nat Methods. 2018 Nov 19. pii: 10.1038/s41592-018-0184-y. doi: 
            10.1038/s41592-018-0184-y.
  PUBMED    30455464
REFERENCE   2  (bases 1 to 5638)
  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 
            Methods. 2018 Nov 19. pii: 10.1038/s41592-018-0184-y. doi: 
            10.1038/s41592-018-0184-y."
COMMENT     Sequence Label: pAAV-TRE-dDIO-vCre
FEATURES             Location/Qualifiers
     source          1..5638
                     /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        148..462
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    156..174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    192..210
                     /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    334..352
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    370..388
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     435..459
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    473..504
                     /label=rox
                     /bound_moiety="Dre recombinase"
     protein_bind    1700..1731
                     /label=rox
                     /bound_moiety="Dre recombinase"
     misc_feature    1778..2366
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1831..1851)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2249..2260)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2385..2861
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2584..2603)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2901..3041
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2901..3030
                     /label=AAV2 ITR
     rep_origin      3116..3571
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3203..3222)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3413..3434
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3588..3607)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3707..3729
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3767..3785)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3853..3957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3958..4026,4027..4818)
                     /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(4176..4195)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4989..5577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5478..5497
                     /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 gcgtctcgag tttactccct atcagtgata gagaacgtat
      181 gtcgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc tatcagtgat
      241 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc
      301 cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga gaacgtatgt
      361 cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt acggtgggag
      421 gcctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagaat tctaacttta
      481 aataatgcca attatttaaa gttagctaac tttaaataat gtcgattatt taaagttagg
      541 cgcgccttag tccttttcgt cctttcccca ctgcattggt gctgaccgag tcttgaccct
      601 gctatgtgcc attgctccgt ctcgagccag aatggctcgt ccatagcgtg tcaccatagc
      661 cccactagac cacctgccgg actgctggat ctgcagagta ttgtatcctt tcttcagcag
      721 gtcctgtgct gctccgactc tggcgctatg tgctgtaaaa ggcttcacgc cctgccgtcc
      781 cagatccagt gtttcccagg ctgaatagaa gaccccctcc acggtttttg tggtaatagg
      841 cttatgcagc acctcgccag tctgtttatc tttcttgggc ttgatacatg tattgtgttt
      901 gctgactccc acaaacagga acccatcgga gctcatgtcc agcttgccgg cttcggtgta
      961 gtccatcagc agtgagacca catcctgact caggatgcaa gtgtctggct ctccagaatg
     1021 gttggtctta gtaataggga ttgtcagaat agcggtccca tcgccggcca gctccatgtc
     1081 ggagacgcgg atatttgcca gttcagaagc tcgcagcatg gactcataag ccacggccag
     1141 cagggccagg tttcgcctca gcagcagttt gtcgcttctg taccacagtg atgtcagctt
     1201 cagcaggtgc tgttcgttga aggggctggc ctgtttgact gcttcgccct ctctcacttt
     1261 cttccgtgca atagccttca gtctatcctc cacatagata tccagacaag ggtcggggca
     1321 tccagcgacc ctatgcactc tggaaatggc ccagcggtac acgctcaggg tattgcggtg
     1381 cagttcctcg gccctttcga taaagaatgc ctcgacagtg tcgggtgttg ctggcagact
     1441 atgcctgtgg ttagccagac accaatccac aaagcaggtc caagcactct ccagtctccg
     1501 cagagtattt ggagagtacc gttcctcccg cttcttgaac tcgttcagca ggtgctcaaa
     1561 agcggctttg aactgttcat tctctgccac gttgatgccc ttcttctgtg cggcctgaat
     1621 ggcggtcttg acatcatcgg gtcggactcc tgaaaaatct cccagcagtg acagctggtt
     1681 ctcaatcatg gtggctagct aactttaaat aattggcatt atttaaagtt agctaacttt
     1741 aaataatcga cattatttaa agttaaagct tatcgataat caacctctgg attacaaaat
     1801 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1861 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     1921 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     1981 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2041 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2101 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2161 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     2221 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2281 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     2341 gatctccctt tgggccgcct ccccgcatcg ataccgagat ctacgggtgg catccctgtg
     2401 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     2461 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     2521 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     2581 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     2641 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     2701 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     2761 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     2821 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     2881 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2941 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3001 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3061 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3121 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3181 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3241 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3301 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3361 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3421 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3481 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3541 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3601 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3661 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3721 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3781 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3841 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3901 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3961 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4021 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4081 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4141 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4201 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4261 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4321 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4381 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4441 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4501 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4561 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4621 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4681 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4741 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4801 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4861 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4921 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4981 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5041 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5101 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5161 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5221 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5281 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5341 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5401 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5461 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5521 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5581 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
