LOCUS       sequence-377887-        6443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FlpO dependent TdTomato expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  AUTHORS   Winnubst J, Bas E, Ferreira TA, Wu Z, Economo MN, Edson P, Arthur 
            BJ, Bruns C, Rokicki K, Schauder D, Olbris DJ, Murphy SD, Ackerman 
            DG, Arshadi C, Baldwin P, Blake R, Elsayed A, Hasan M, Ramirez D, 
            Dos Santos B, Weldon M, Zafar A, Dudman JT, Gerfen CR, Hantman AW, 
            Korff W, Sternson SM, Spruston N, Svoboda K, Chandrashekar J
  TITLE     Reconstruction of 1,000 Projection Neurons Reveals New Cell Types 
            and Organization of Long-Range Connectivity in the Mouse Brain.
  JOURNAL   Cell. 2019 Sep 19;179(1):268-281.e13. doi: 
            10.1016/j.cell.2019.07.042. Epub 2019 Sep 5.
  PUBMED    31495573
REFERENCE   2  (bases 1 to 6443)
  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: 
            "10.1016/j.cell.2019.07.042"; journalName: "Cell"; date: 
            "2019-09-19- 19"; volume: "179"; issue: "1"; pages: "268-281.e13"
COMMENT     Sequence Label: pAAV-CAG-FRT-Rev-TdTomato
FEATURES             Location/Qualifiers
     source          1..6443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(97..116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      910..1498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1399..1418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1735..1864
                     /label=AAV2 ITR
     polyA_signal    2026..2147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2063..2082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2117..2136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2190..2778
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2296..2307
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2705..2725
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    2813..2846
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    2875..2922
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2942..2951
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4401..4434)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    complement(4477..4524)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(4565..4584)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4646..4668)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     repeat_region   5535..5664
                     /label=AAV2 ITR
     rep_origin      5736..6191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5823..5842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6033..6054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6217..6321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6322..6390,6391..6443,1..739)
                     /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"
ORIGIN
        1 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
       61 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      121 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      181 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      241 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      301 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      361 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      421 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      481 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      541 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      601 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      661 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      721 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      781 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      841 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      901 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      961 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1021 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1081 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1141 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1201 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1261 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1321 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1381 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1441 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1501 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1561 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1621 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     1681 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcgc
     1741 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
     1801 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     1861 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag
     1921 atccttatcg attttaccac atttgtagag gttttacttg ctttaaaaaa cctcccacat
     1981 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2041 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2101 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg
     2161 aagcggccgc ccgggtcgac ggtatcgatg cggggaggcg gcccaaaggg agatccgact
     2221 cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca ggccgcggga
     2281 aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat
     2341 ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg acaacaccac
     2401 ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgat
     2461 gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga gctgacaggt
     2521 ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca ccacgccacg
     2581 ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt tatacaagga
     2641 ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta
     2701 tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt
     2761 gtaatccaga ggttgattat cgataagctt cctgcaggga agttcctatt ccgaagttcc
     2821 tattcttcaa atagtatagg aacttcttgg atcaaaatag gagcgatcgc aatggaagtt
     2881 cctattccga agttcctatt ctctagaaag tataggaact tcgtttaaac tggccggcct
     2941 cgccaccatg gttagcaagg gtgaagaggt gattaaagag tttatgcgtt ttaaagtacg
     3001 catggagggt agcatgaacg gtcatgagtt tgagatcgag ggcgaaggcg aaggtcgtcc
     3061 gtacgagggt acgcagaccg ctaaactgaa agtaaccaaa ggcggtcctc tgccgttcgc
     3121 gtgggacatc ctgtccccgc agtttatgta tggttccaag gcctacgtca aacacccagc
     3181 cgatatcccg gactacaaga agctgtcttt cccggagggt ttcaagtggg agcgtgtaat
     3241 gaactttgag gacggtggcc tggttaccgt cacccaggat agctctctgc aagatggcac
     3301 cctgatctac aaggtgaaga tgcgtggtac gaactttccg ccggatggcc cggtgatgca
     3361 gaaaaagacg atgggatggg aagcatctac cgagcgcctg tatccgcgtg acggtgttct
     3421 gaaaggtgag atccaccagg cgctgaaact gaaggacggc ggtcactacc tggttgaatt
     3481 taagacgatc tatatggcta aaaagccggt tcaactgcca ggttactact acgtggacac
     3541 caaactggat atcacgtctc acaacgaaga ttataccatc gtcgaacagt atgaacgtag
     3601 cgagggccgt caccacctgt tcctgggcca cggtactggt tctacgggtt ctggttcctc
     3661 tggcactgcg tcttccgaag acaacaacat ggcggtgatc aaagagttca tgaggttcaa
     3721 agtacgtatg gaaggttcta tgaacggcca tgagttcgag attgaaggtg agggcgaggg
     3781 tcgcccatac gagggcactc agaccgcaaa gctgaaagtt accaaaggtg gcccactgcc
     3841 ttttgcgtgg gatatcctgt ctccgcagtt catgtacggt tccaaagcct acgtaaaaca
     3901 cccggcagac atcccggatt acaaaaagct gtccttccca gagggcttca aatgggaacg
     3961 tgttatgaat ttcgaagatg gcggcctggt cactgttacg caggacagca gcctgcaaga
     4021 cggtactctg atttacaaag tgaaaatgcg tggcactaac ttcccgcctg acggtccagt
     4081 aatgcagaag aaaactatgg gctgggaagc tagcaccgaa cgtctgtacc cgcgcgacgg
     4141 tgtgctgaaa ggcgagattc accaggccct gaagctgaag gatggtggtc actatctggt
     4201 tgagttcaaa actatctaca tggcaaaaaa gcctgtacaa ctgccgggct attactacgt
     4261 tgacactaaa ctggacatca cctcccacaa cgaggattat actattgttg aacagtacga
     4321 gcgttctgaa ggtcgccatc atctgtttct gtatggtatg gacgaactgt ataaataaac
     4381 gcgtttaatt aagtttaaac gaagttccta tactatttga agaataggaa cttcggaata
     4441 ggaacttcag agcgctttgg ccgctccggc caaagggaag ttcctatact ttctagagaa
     4501 taggaacttc ggaataggaa cttcggcgcg ccggatccaa ttctttgcca aaatgatgag
     4561 acagcacaat aaccagcacg ttgcccagga gctgtaggaa aaagaagaag gcatgaacat
     4621 ggttagcaga ggctctagag ccgccggtca cacgccagaa gccgaacccc gccctgcccc
     4681 gtcccccccg aaggcagccg tccccccgcg gacagccccg agccgctccc cccgcacaaa
     4741 gggccctccc ggagcccctc aaggctttca cgcagccaca gaaaagaaac aagccgtcat
     4801 taaaccaagc gctaattaca gcccggagga gaagggccgt cccgcccgct cacctgtggg
     4861 agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg cggcggcgga gcggggcacg
     4921 gggcgaaggc agcgcgcagc gactcccgcc cgccgcgcgc ttcgcttttt atagggccgc
     4981 cgccgccgcc gcctcgccat aaaaggaaac tttcggagcg cgccgctctg attggctgcc
     5041 gccgcmcctc yccgccycsc cccsccccsc cccycscccc sccccccccc sccyggcgcg
     5101 cscccccccc ccccccgccc ccatcgctgc acaaaataat taaaaaataa ataaatacaa
     5161 aattgggggt ggggaggggg gggagatggg gagagtgaag cagaacgtgg ggctcacctc
     5221 gaccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     5281 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     5341 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     5401 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     5461 tgacgtcaat ggtacgatct tcctagagca tggctacgta gataagtagc atggcgggtt
     5521 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     5581 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     5641 ctcagtgagc gagcgagcgc gcagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5701 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     5761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5941 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6001 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6061 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6181 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6241 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6301 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6361 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6421 gctgaagatc agttgggtgc acg
//
