LOCUS       sequence-183001-        7402 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 7402)
  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 7402)
  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..7402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        234..537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        539..816
                     /label=chicken beta-actin promoter
     intron          817..1833
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1757..1779
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1841..1860
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1961..1994)
                     /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    2162..2296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2416..2550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2670..2804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2833..2866)
                     /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      2940..2949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3079..3097)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3347..3366
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3541..3564
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4429..4446)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4435..4642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4665..4805
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4665..4794
                     /label=AAV2 ITR
     rep_origin      4880..5335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4967..4986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5177..5198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5352..5371)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5471..5493
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5531..5549)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5617..5721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5722..5790,5791..6582)
                     /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(5940..5959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6753..7341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7242..7261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgaaacaa ttctgcagga atctagttat
      181 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
      541 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
      601 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
      661 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      721 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      781 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      841 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      901 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      961 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
     1021 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
     1081 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     1141 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1201 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1261 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     1321 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     1381 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc
     1441 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1501 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1561 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1621 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1681 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1741 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1801 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1861 ctgtctcatc attttggcaa agaattgatt tgataccgcg ggccccgcgt ggacggtacc
     1921 cttcaggtcg acatttatcc ctaaacagga taacactagc ataacttcgt ataggatact
     1981 ttatacgaag ttattcgcga tgaataaatg aaagcttgca gatctgcgac tctagaggat
     2041 ctgcgactct agaggatcat aatcagccat accacatttt gtagaggttt tacttgcttt
     2101 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2161 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2221 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2281 ttatcatgtc tggatctgcg actctagagg atcataatca gccataccac atttgtagag
     2341 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2401 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2461 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2521 ctcatcaatg tatcttatca tgtctggatc tgcgactcta gaggatcata atcagccata
     2581 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2641 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2701 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2761 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccccatc aagctgatcc
     2821 ggaaccctta atataacttc gtataggata ctttatacga agttattagc tagttacagt
     2881 accggcttag ttaggaattc taaatttcga aagtcgacgt cggttggcgc gccgctagcg
     2941 ccaccatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca
     3001 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3061 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     3121 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg
     3181 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     3241 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc
     3301 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
     3361 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     3421 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     3481 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca
     3541 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
     3601 agggccgcca ccacctgttc ctggggcatg gtaccggcag caccggcagc ggcagctccg
     3661 gcaccgcctc ctccgaggac aacaacatgg ccgtgattaa ggagtttatg aggtttaaag
     3721 tcaggatgga gggcagcatg aacgggcacg agtttgagat tgaaggagag ggggaaggga
     3781 ggccctatga gggcacccag acagcaaagc tgaaagtgac aaagggcggc cctctgccat
     3841 tcgcatggga catcctgtct cctcagttta tgtatggaag taaagcctac gtgaaacacc
     3901 ctgctgacat tccagattac aagaaactga gcttcccaga ggggtttaag tgggaaagag
     3961 tgatgaactt cgaagacggg ggcctggtga ctgtcaccca ggacagctct ctgcaggacg
     4021 gcactctgat ctataaggtg aaaatgcgcg gcaccaattt tccccctgat ggcccagtca
     4081 tgcagaagaa gacaatgggc tgggaggctt caacagaaag gctgtacccc agagacggcg
     4141 tgctgaaggg agagatccac caggcactga agctgaaaga tggagggcat tatctggtcg
     4201 aattcaaaac catctacatg gccaagaaac ccgtgcagct gcctggctac tattacgtcg
     4261 acacaaagct ggatattact agccataacg aagactacac cattgtggag cagtatgagc
     4321 ggagcgaggg gaggcaccat ctgtttctgt acggcatgga cgagctgtac aagtaagcta
     4381 gcggcgcgcc aagggtttaa accccctggc cggccttgcg aagatctgcc tcgactgtgc
     4441 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4501 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4561 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4621 acaatagcag gcatgctggg gacacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     4681 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4741 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     4801 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4861 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4921 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4981 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5041 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     5101 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5161 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5221 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5281 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     5341 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5401 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5461 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5521 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5581 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5641 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5701 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5761 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5821 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5881 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5941 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6001 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6061 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6121 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6181 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6241 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6301 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6361 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6421 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6481 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6541 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6601 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6661 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6721 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6781 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6841 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6901 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6961 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7021 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7081 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7141 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7201 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7261 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7321 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7381 tgctggcctt ttgctcacat gt
//
