LOCUS       sequence-243967-        6931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flpo under the CAG promoter in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6931)
  AUTHORS   Fujimoto S, Leiwe MN, Aihara S, Sakaguchi R, Muroyama Y, Kobayakawa 
            R, Kobayakawa K, Saito T, Imai T
  TITLE     Activity-dependent local protection and lateral inhibition control 
            synaptic competition in developing mitral cells in mice.
  JOURNAL   Dev Cell. 2023 Jul 24;58(14):1221-1236.e7. doi: 
            10.1016/j.devcel.2023.05.004. Epub 2023 Jun 7.
  PUBMED    37290446
REFERENCE   2  (bases 1 to 6931)
  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.devcel.2023.05.004"; journalName: "Dev Cell"; date: 
            "2023-07-24- 24"; volume: "58"; issue: "14"; pages: "1221-1236.e7"
COMMENT     Sequence Label: pCAG-Flpo
FEATURES             Location/Qualifiers
     source          1..6931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          165..1181
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1105..1127
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1189..1208
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        1240..1739
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1275..1298)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1672..1691
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1749..1758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1755..1760,1761..1781,1782..3053)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     primer_bind     complement(3078..3095)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3084..3308
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3456..3475)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3521..3576
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3575..3594)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3937..3953)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3937..3953)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3950..3972)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3961..3977
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3985..3991,3992..4009,4010..4015))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4030..4051
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4110..4305
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4156..4291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4311..4445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4348..4367)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4402..4421
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4513..4530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4684..5272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4764..4783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5443..6234,6235..6303))
                     /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     6066..6085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6304..6408)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6439..6818
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6820..6931,1..164)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
       61 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      121 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
      181 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      241 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      301 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
      361 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
      421 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
      481 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
      541 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
      601 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
      661 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
      721 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
      781 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
      841 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
      901 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
      961 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     1021 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     1081 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1141 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1201 ttattgtgct gtctcatcat tttggcaaag aattctaccg ggtaggggag gcgcttttcc
     1261 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     1321 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     1381 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     1441 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     1501 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     1561 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     1621 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     1681 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     1741 ccatcgccgc caccatggct cctaagaaga agaggaaggt gatgagccag ttcgacatcc
     1801 tgtgcaagac cccccccaag gtgctggtgc ggcagttcgt ggagagattc gagaggccca
     1861 gcggcgagaa gatcgccagc tgtgccgccg agctgaccta cctgtgctgg atgatcaccc
     1921 acaacggcac cgccatcaag agggccacct tcatgagcta caacaccatc atcagcaaca
     1981 gcctgagctt cgacatcgtg aacaagagcc tgcagttcaa gtacaagacc cagaaggcca
     2041 ccatcctgga ggccagcctg aagaagctga tccccgcctg ggagttcacc atcatccctt
     2101 acaacggcca gaagcaccag agcgacatca ccgacatcgt gtccagcctg cagctgcagt
     2161 tcgagagcag cgaggaggcc gacaagggca acagccacag caagaagatg ctgaaggccc
     2221 tgctgtccga gggcgagagc atctgggaga tcaccgagaa gatcctgaac agcttcgagt
     2281 acaccagcag gttcaccaag accaagaccc tgtaccagtt cctgttcctg gccacattca
     2341 tcaactgcgg caggttcagc gacatcaaga acgtggaccc caagagcttc aagctggtgc
     2401 agaacaagta cctgggcgtg atcattcagt gcctggtgac cgagaccaag acaagcgtgt
     2461 ccaggcacat ctactttttc agcgccagag gcaggatcga ccccctggtg tacctggacg
     2521 agttcctgag gaacagcgag cccgtgctga agagagtgaa caggaccggc aacagcagca
     2581 gcaacaagca ggagtaccag ctgctgaagg acaacctggt gcgcagctac aacaaggccc
     2641 tgaagaagaa cgccccctac cccatcttcg ctatcaagaa cggccctaag agccacatcg
     2701 gcaggcacct gatgaccagc tttctgagca tgaagggcct gaccgagctg acaaacgtgg
     2761 tgggcaactg gagcgacaag agggcctccg ccgtggccag gaccacctac acccaccaga
     2821 tcaccgccat ccccgaccac tacttcgccc tggtgtccag gtactacgcc tacgacccca
     2881 tcagcaagga gatgatcgcc ctgaaggacg agaccaaccc catcgaggag tggcagcaca
     2941 tcgagcagct gaagggcagc gccgagggca gcatcagata ccccgcctgg aacggcatca
     3001 tcagccagga ggtgctggac tacctgagca gctacatcaa caggcggatc tgatgagatc
     3061 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3121 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3181 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3241 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3301 ctctatggct tctgaggcgg aaagaaccag ctggggctcg agatccacta gttctagcct
     3361 cgaggctaga gcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     3421 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     3481 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     3541 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     3601 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     3661 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     3721 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     3781 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     3841 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     3901 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3961 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4021 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4081 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     4141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     4321 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4381 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4441 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4501 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4561 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4621 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4681 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4741 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4801 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4861 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4921 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4981 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5041 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5101 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5161 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5221 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5281 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5341 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5401 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5461 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5521 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5581 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5641 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5701 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5761 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5821 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5881 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5941 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6001 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6061 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6121 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6181 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6241 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6301 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6361 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6421 aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg
     6481 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6541 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6601 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6661 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6721 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6781 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
     6841 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
     6901 aattattttg tgcagcgatg ggggcggggg g
//
