LOCUS       sequence-193827-        7390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV mediated Cre-dependent expression of tdtomato (Lox-Stop-Lox).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7390)
  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 7390)
  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: 
            "10.1038/nature13186"; journalName: "Nature"; date: "2014-04-10- 
            10"; volume: "508"; issue: "7495"; pages: "207-14"
COMMENT     Sequence Label: pAAV.CAG.LSL.tdTomato
FEATURES             Location/Qualifiers
     source          1..7390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..147
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   18..147
                     /label=AAV2 ITR
     enhancer        240..543
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        545..821
                     /label=chicken beta-actin promoter
     intron          822..1838
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1762..1784
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1846..1865
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1966..1999)
                     /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)."
     polyA_signal    2167..2301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2421..2555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2675..2809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2838..2871)
                     /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)."
     regulatory      2945..2954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3084..3102)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3352..3371
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3546..3569
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4434..4451)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4440..4647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4670..4799
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4861..5449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4941..4960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5620..6411,6412..6480))
                     /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     6243..6262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6481..6585)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6653..6671
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6709..6731)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6831..6850
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6867..7322)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7004..7025)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7216..7235
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
       61 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
      121 tggccaactc catcactagg ggttcctgcg gccgcacgcg aaacaattct gcaggaatct
      181 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      361 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      541 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
      601 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
      661 gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
      721 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
      781 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      841 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      901 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
      961 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
     1021 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
     1081 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
     1141 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
     1201 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     1261 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
     1321 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
     1381 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     1441 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
     1501 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     1561 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     1621 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     1681 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     1741 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     1801 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     1861 ttgtgctgtc tcatcatttt ggcaaagaat tgatttgata ccgcgggccc cgcgtggacg
     1921 gtacccttca ggtcgacatt tatccctaaa caggataaca ctagcataac ttcgtatagg
     1981 atactttata cgaagttatt cgcgatgaat aaatgaaagc ttgcagatct gcgactctag
     2041 aggatctgcg actctagagg atcataatca gccataccac attttgtaga ggttttactt
     2101 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2161 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2221 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2281 gtatcttatc atgtctggat ctgcgactct agaggatcat aatcagccat accacatttg
     2341 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2401 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2521 ccaaactcat caatgtatct tatcatgtct ggatctgcga ctctagagga tcataatcag
     2581 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2641 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2701 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2761 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc ccatcaagct
     2821 gatccggaac ccttaatata acttcgtata ggatacttta tacgaagtta ttagctagtt
     2881 acagtaccgg cttagttagg aattctaaat ttcgaaagtc gacgtcggtt ggcgcgccgc
     2941 tagcgccacc atggtgagca agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt
     3001 gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3061 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt
     3121 cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc
     3181 cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3241 gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg
     3301 cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat
     3361 gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt
     3421 gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga
     3481 gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga
     3541 caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg
     3601 ctccgagggc cgccaccacc tgttcctggg gcatggtacc ggcagcaccg gcagcggcag
     3661 ctccggcacc gcctcctccg aggacaacaa catggccgtg attaaggagt ttatgaggtt
     3721 taaagtcagg atggagggca gcatgaacgg gcacgagttt gagattgaag gagaggggga
     3781 agggaggccc tatgagggca cccagacagc aaagctgaaa gtgacaaagg gcggccctct
     3841 gccattcgca tgggacatcc tgtctcctca gtttatgtat ggaagtaaag cctacgtgaa
     3901 acaccctgct gacattccag attacaagaa actgagcttc ccagaggggt ttaagtggga
     3961 aagagtgatg aacttcgaag acgggggcct ggtgactgtc acccaggaca gctctctgca
     4021 ggacggcact ctgatctata aggtgaaaat gcgcggcacc aattttcccc ctgatggccc
     4081 agtcatgcag aagaagacaa tgggctggga ggcttcaaca gaaaggctgt accccagaga
     4141 cggcgtgctg aagggagaga tccaccaggc actgaagctg aaagatggag ggcattatct
     4201 ggtcgaattc aaaaccatct acatggccaa gaaacccgtg cagctgcctg gctactatta
     4261 cgtcgacaca aagctggata ttactagcca taacgaagac tacaccattg tggagcagta
     4321 tgagcggagc gaggggaggc accatctgtt tctgtacggc atggacgagc tgtacaagta
     4381 agctagcggc gcgccaaggg tttaaacccc ctggccggcc ttgcgaagat ctgcctcgac
     4441 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     4501 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4561 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4621 ggaagacaat agcaggcatg ctggggacac gtgcggaccg agcggccgca ggaaccccta
     4681 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4741 aaggtcgccc gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga
     4801 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4861 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4921 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4981 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5041 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5101 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5161 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5221 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5281 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5341 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5401 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5461 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5521 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5581 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5641 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5701 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5761 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5821 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5881 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5941 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6001 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6061 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6121 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6181 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6241 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6301 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6361 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6421 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6481 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6541 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6601 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6661 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6721 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6781 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     6841 gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc
     6901 gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc
     6961 ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag
     7021 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     7081 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     7141 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     7201 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt
     7261 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     7321 gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     7381 taccgcatca
//
