LOCUS       sequence-487260-        7207 bp    DNA     circular SYN 26-SEP-2025
DEFINITION  Can be used to generate AAV virus that will express tdTomato in the 
            presence of Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7207)
  AUTHORS   Oh SW, Harris JA, Ng L, Winslow B, Cain N, Mihalas S, Wang Q, Lau C,
            Kuan L, Henry AM, Mortrud MT, Ouellette B, Nguyen TN, Sorensen SA, 
            Slaughterbeck CR, Wakeman W, Li Y, Feng D, Ho A, Nicholas E, 
            Hirokawa KE, Bohn P, Joines KM, Peng H, Hawrylycz MJ, Phillips JW, 
            Hohmann JG, Wohnoutka P, Gerfen CR, Koch C, Bernard A, Dang C, Jones
            AR, Zeng H
  TITLE     A mesoscale connectome of the mouse brain.
  JOURNAL   Nature. 2014 Apr 10;508(7495):207-14. doi: 10.1038/nature13186. Epub
            2014 Apr 2.
  PUBMED    24695228
REFERENCE   2  (bases 1 to 7207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature13186"; journalName: "Nature"; date: "2014-04-10- 
            10"; volume: "508"; issue: "7495"; pages: "207-14"
COMMENT     Sequence Label: AAV pCAG-FLEX-tdTomato-WPRE
FEATURES             Location/Qualifiers
     source          1..7207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        223..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          829..1845
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1769..1791
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1853..1872
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1949..1982)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1993..2026)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(2855..2878)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3053..3072)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3322..3340
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3470..3479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3510..3543
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3554..3587
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    3611..4199
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3664..3684)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4082..4093)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4234..4251)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4240..4447
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4470..4610
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4470..4599
                     /label=AAV2 ITR
     rep_origin      4685..5140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4772..4791)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4982..5003
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5157..5176)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5276..5298
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5336..5354)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5422..5526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5527..5595,5596..6387)
                     /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(5745..5764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6558..7146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7047..7066
                     /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 gcgaaacaat tctgcaggaa tctagttatt aatagtaatc
      181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      601 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggg ggcggccggg
      661 gggggggggg gggggggggg gggggggggg gggggggggg gggggcgggg cggggcgagg
      721 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      781 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      841 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      901 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      961 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
     1021 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
     1081 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     1141 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     1201 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     1261 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     1321 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     1381 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     1441 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     1501 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     1561 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     1621 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     1681 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     1741 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     1801 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     1861 ctggttattg tgctgtctca tcattttggc aaagaattga tttgataccg cgggccccgc
     1921 gtggacggta cccttcaggt cgacgtcgat aacttcgtat agcatacatt atacgaagtt
     1981 atccgaatcg caataacttc gtataggata ctttatacga agttatttgg cgcgccgcta
     2041 gcttacttgt acagctcgtc catgccgtac agaaacagat ggtgcctccc ctcgctccgc
     2101 tcatactgct ccacaatggt gtagtcttcg ttatggctag taatatccag ctttgtgtcg
     2161 acgtaatagt agccaggcag ctgcacgggt ttcttggcca tgtagatggt tttgaattcg
     2221 accagataat gccctccatc tttcagcttc agtgcctggt ggatctctcc cttcagcacg
     2281 ccgtctctgg ggtacagcct ttctgttgaa gcctcccagc ccattgtctt cttctgcatg
     2341 actgggccat cagggggaaa attggtgccg cgcattttca ccttatagat cagagtgccg
     2401 tcctgcagag agctgtcctg ggtgacagtc accaggcccc cgtcttcgaa gttcatcact
     2461 ctttcccact taaacccctc tgggaagctc agtttcttgt aatctggaat gtcagcaggg
     2521 tgtttcacgt aggctttact tccatacata aactgaggag acaggatgtc ccatgcgaat
     2581 ggcagagggc cgccctttgt cactttcagc tttgctgtct gggtgccctc atagggcctc
     2641 ccttccccct ctccttcaat ctcaaactcg tgcccgttca tgctgccctc catcctgact
     2701 ttaaacctca taaactcctt aatcacggcc atgttgttgt cctcggagga ggcggtgccg
     2761 gagctgccgc tgccggtgct gccggtacca tgccccagga acaggtggtg gcggccctcg
     2821 gagcgctcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccagcttg
     2881 gtgtccacgt agtagtagcc gggcagttgc acgggcttct tggccatgta gatggtcttg
     2941 aactccacca ggtagtggcc gccgtccttc agcttcaggg cctggtggat ctcgcccttc
     3001 agcacgccgt cgcgggggta caggcgctcg gtggaggcct cccagcccat ggtcttcttc
     3061 tgcattacgg ggccgtcggg ggggaagttg gtgccgcgca tcttcacctt gtagatcagc
     3121 gtgccgtcct gcagggagga gtcctgggtc acggtcacca gaccgccgtc ctcgaagttc
     3181 atcacgcgct cccacttgaa gccctcgggg aaggacagct tcttgtaatc ggggatgtcg
     3241 gcggggtgct tcacgtacgc cttggagccg tacatgaact ggggggacag gatgtcccag
     3301 gcgaagggca gggggccgcc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag
     3361 gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga gccctccatg
     3421 cgcaccttga agcgcatgaa ctctttgatg acctcctcgc ccttgctcac catggtggcg
     3481 ctagcggcgc gccaacctgg ccggccttga taacttcgta taatgtatgc tatacgaagt
     3541 tatccgaatc gcaataactt cgtataaagt atcctatacg aagttatcga agatcatcaa
     3601 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3661 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3721 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3781 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3841 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3901 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3961 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     4021 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     4081 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     4141 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     4201 tcgataccga gcgctgctcg agagatcgat ctgcctcgac tgtgccttct agttgccagc
     4261 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4321 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4381 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4441 ctggggacac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     4501 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4561 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     4621 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     4681 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4741 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     4801 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4861 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4921 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4981 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     5041 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     5101 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     5161 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     5221 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5281 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     5341 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5401 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5461 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5521 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5581 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5641 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5701 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5761 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5821 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5881 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5941 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6001 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6061 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6121 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6181 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6241 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6301 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6361 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6421 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6481 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6541 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6601 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6661 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6721 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6781 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6841 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6901 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6961 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7021 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7081 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7141 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7201 cacatgt
//
