LOCUS       sequence-201082-        7498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome encoding expression of the nuclear localized 
            fluorescent protein tdTomato from the MBP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7498)
  AUTHORS   Challis RC, Ravindra Kumar S, Chan KY, Challis C, Beadle K, Jang MJ,
            Kim HM, Rajendran PS, Tompkins JD, Shivkumar K, Deverman BE, 
            Gradinaru V
  TITLE     Systemic AAV vectors for widespread and targeted gene delivery in 
            rodents.
  JOURNAL   Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3.
  PUBMED    30626963
REFERENCE   2  (bases 1 to 7498)
  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"; doi: "10"; journalName: 
            "Nat Protoc"; date: "2019-02"; volume: "14"; issue: "2"; pages: 
            "379-414"
COMMENT     Sequence Label: pAAV-MBP-2xNLS-tdTomato
FEATURES             Location/Qualifiers
     source          1..7498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2101..2110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2116..2136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      2143..2152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2149..3570
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNF
                     EDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLK
                     GEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERS
                     EGRHHLFLYGMDELYK"
     CDS             3577..3597
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3625..4213
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3678..3698)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4096..4107)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4245..4721
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4444..4463)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4761..4901
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4761..4890
                     /label=AAV2 ITR
     rep_origin      4976..5431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5063..5082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5273..5294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5448..5467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5567..5589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5627..5645)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5713..5817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5818..5886,5887..6678)
                     /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(6036..6055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6849..7437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7338..7357
                     /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 gcgttccttc ctgcttaggc cgtgtctgct ctgtcagtct
      181 ttcctctggc tctagtctgt ccatctgcag ggataacctc ccaagggccc tcttgttggc
      241 tgttcacctt ccgtctgtct tcatagcctc ctgatgagag agcgcgcaca cacaggctac
      301 ctgactgatg tgttgctgca ctgtggggtc cagtgacaca gctgtggagg tcaggaaccc
      361 agaattaagc tgtctgggag ccacaagttt caagagtcca tgctatggct cctacatgca
      421 ggtggtgcca gcagcattcc ttggtgtcct gtggctgtgg ttgcatctct gactctcctc
      481 cacctatcac attccttctc cagtaccagg cctctgtgtc tcagttttcc tctctcacaa
      541 gatccctagc ttcaggattg cacccacctc cactagccat gagcccgtcc tcactggatc
      601 acgtctgcaa agactctatt tccagttaaa gtcacagaca cgggcataga gagggggata
      661 ggatatgaaa atatcagttg ggggaggggc attcaattct tagctcacat tttcttcatc
      721 caatttcccc cactagaatg taagctttga gagaaaaggg accagatctt attcctcacc
      781 gtggctttaa cacttagaga aaatgcatcc cctctaatca ataagtcatc gacagtgggt
      841 agatggagga acggcagtgc gtagtaggat gcgtgcaagc atagtctcgt gcatgggtgc
      901 atagatcgct gggcaggtgg acaaggtggg ggtggataaa gaagtgggta gatgattgat
      961 gttaggtaaa tatcactggg tggacagatg ggtggtaggt ggatggatgg ttagaatagt
     1021 cagaagaggg atggattgat aaggtgaaca gatgataaat gggtgataga ctggaagggt
     1081 tgtcaaaaga ggataaggga agtgtgagct agccgtattt ctaaggtcag taatagagtt
     1141 gggagaagag gttaagttac atccatttaa acctcacacg aagctgagag ggaatggact
     1201 tgctgccgtt ggtgaggaaa gcgttgcatt tcccgtgtgc ttggttgtga agtgctcagg
     1261 tcccacatga agcagtcagg ttactgcggc ttacagagga gccagatcca aatgccccga
     1321 gtaagcacgt ccccgagcca gaggcctcca gcggaatccg ggagagggat tgctcagtgc
     1381 cctgcttccc tggactgtaa gctgcagaaa gatgtgggaa gtcctgttct ccactgagaa
     1441 cactaaaagc accttttgtc aaacgaccgc ttcacatctg gggcttgtgc actggtggcc
     1501 ttttaaacca gagacaaccc acaagatacc taacctgcgg ggctctctgg tacagtgagc
     1561 aactcaggaa atgctttggc ttgattgctg tgggctctca ggccatcgcc ctctggagtg
     1621 gttcttttaa tgagaacctg aagattggcc cctgagccat gtataccaag caagctcaat
     1681 ccaggttagc tccctctggt tggggcaagc taacgtgctc cttgggcccc gcgcgtaact
     1741 gtgcgtttta taggagacag ctagttcaag accccaggaa gaaagcggct ttgtccccct
     1801 ctaggcctcg tacaggccca cattcatatc tcattgttgt tgcaggggag gcagatgcga
     1861 tccagaacaa tgggacctcg gctgaggaca cggcggtgac agactccaag cacacagcag
     1921 acccaaagaa taactggcaa ggcgcccacc cagctgaccc agggaaccgc ccccacttga
     1981 tccgcctctt ttcccgagat gccccgggaa gggaggacaa caccttcaaa gacaggccct
     2041 cagagtccga cgagcttcag accatccaag aagaccccac agcagcttcc ggagggtacc
     2101 gccaccatgg gaagcccaaa gaagaagcgt aaggtaggat ccgccaccat ggtgagcaag
     2161 ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac
     2221 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2281 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     2341 cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag
     2401 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt
     2461 ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta caaggttaag
     2521 atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg
     2581 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag
     2641 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc
     2701 aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc
     2761 cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg
     2821 ttcctggggc atggcaccgg cagcaccggc agcggcagct ccggcaccgc ctcctccgag
     2881 gacaacaaca tggccgtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc
     2941 atgaacggcc acgagttcga gatcgagggc gagggccgcc cctacgaggg cacccagacc
     3001 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     3061 cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag
     3121 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt
     3181 ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta caaggttaag
     3241 atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg
     3301 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag
     3361 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc
     3421 aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc
     3481 cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg
     3541 ttcctgtacg gcatggacga gctgtacaag ggaagcccca agaaaaagcg gaaggtgtga
     3601 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3661 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3721 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3781 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3841 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3901 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3961 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4021 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     4081 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4141 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4201 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     4261 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     4321 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     4381 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     4441 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     4501 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     4561 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     4621 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     4681 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     4741 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     4801 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4861 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     4921 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4981 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5041 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5101 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5161 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     5221 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5281 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     5341 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5401 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     5461 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5521 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5581 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5641 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5701 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5761 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5821 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5881 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5941 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6001 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6061 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6121 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6181 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6241 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6301 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6361 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6421 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6481 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6541 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6601 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6661 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6721 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6781 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6841 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6901 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6961 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7021 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7081 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7141 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7201 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7261 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7321 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7381 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7441 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
