LOCUS       sequence-297944-        5201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector based on the LiOn integration-coupled translational switch 
            and devoid of TTAA sequences, expressing the fluorescent protein 
            mRFP1 from a CAG promoter upon action of the piggyBac transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5201)
  AUTHORS   Kumamoto T, Maurinot F, Barry-Martinet R, Vaslin C, 
            Vandormael-Pournin S, Le M, Lerat M, Niculescu D, Cohen-Tannoudji M,
            Rebsam A, Loulier K, Nedelec S, Tozer S, Livet J
  TITLE     Direct Readout of Neural Stem Cell Transgenesis with an 
            Integration-Coupled Gene Expression Switch.
  JOURNAL   Neuron. 2020 Aug 19;107(4):617-630.e6. doi: 
            10.1016/j.neuron.2020.05.038. Epub 2020 Jun 18.
  PUBMED    32559415
REFERENCE   2  (bases 1 to 5201)
  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.neuron.2020.05.038"; journalName: "Neuron"; date: 
            "2020-08-19- 19"; volume: "107"; issue: "4"; pages: "617-630.e6"
COMMENT     Sequence Label: LiOn*-CAG∞RFP
FEATURES             Location/Qualifiers
     source          1..5201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          159..1167
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1091..1113
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1232..1241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   1255..1289
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1620..1643)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1818..1837)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2087..2105
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     repeat_region   2223..2285
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     2639..2658
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2657..2712
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2758..2777
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     rep_origin      complement(2924..3512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3004..3023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3683..4474,4475..4543))
                     /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     4306..4325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4544..4648)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4702..5081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5084..5201,1..158)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
       61 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
      121 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg ttgccttcgc
      181 cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      241 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
      301 attgacggct cgtttctttt ctgtggctgc gtgaaagcct tatagggctc cgggagggcc
      361 ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
      421 gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
      481 cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggc
      541 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
      601 ggcgcggcgg tcgggctgta acccccccct gcacccccct ccccgagttg ctgagcacgg
      661 cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg tgccgggcgg
      721 ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc
      781 gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
      841 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgg
      901 cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg cgaagcggtg
      961 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
     1021 ccttctccat ctccagcctc ggggctgccg cagggggacg gctgccttcg ggggggacgg
     1081 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     1141 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttgt tgtgctgtct
     1201 catcattttg gcaaagcacg tgaggaccgg tgccaccatg gcctcctccc ttaaccctag
     1261 aaagatagtc tgcgtaaaat tgacgcatgc atcaattgta actgcggttt atactcccca
     1321 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     1381 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     1441 acagtgagag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     1501 ggctggcaac tagaaggcac taaagtcgga gaattcattc ccgggggaaa aagatcttta
     1561 ggcgccggtg gagtggcggc cctcggcgcg ctcgtactgt tccacgatgg tgtagtcctc
     1621 gttgtgggag gtgatgtcca gcttgatgtc ggtcttgtag gcgccgggca gctgcacggg
     1681 cttcttggcc atgtaggtgg tcttgacctc ggcgtcgtag tggccgccgt ccttcagctt
     1741 cagcctcatc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggtgga
     1801 ggcctcccag cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc
     1861 gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt
     1921 caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga
     1981 cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtactg
     2041 gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccgcccttgg tcaccttcag
     2101 cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc
     2161 gtggccgttc acggagccct ccatgcgcac cttgaagcgc atgaactcct tgatgacatt
     2221 aaccctagaa agataatcat attgtgacgt acgttaaaga taatcatgcg taaaattgac
     2281 gcatgtccgg agccgtagat atcggaaaga aagcggccgc tgcaggtcga gggatcttca
     2341 taagagaaga gggacagcta tgactgggag tagtcaggag aggaggaaaa atctggctag
     2401 taaaacatgt aaggaaaatt ttagggatgt tatagaaaaa aataacacaa aacaaaatat
     2461 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
     2521 ctgtttcata tactgatgac ctctttatag ccacctttgt tcatggcagc cagcatatgg
     2581 catatgttgc caaactctaa accaaatact cattctgatg ttttatatga tttgccctcc
     2641 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     2701 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     2761 tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc acaccagcca
     2821 ccaccttctg ataggcagcc tgcacctgag gagtgaatta gaagctttga gcaaaaggcc
     2881 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2941 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3001 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3061 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3121 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3181 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3241 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3301 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3361 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3421 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3481 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3541 ctgacgctca gtggaacgaa aactcacgtt tagggatttt ggtcatgaga ttatcaaaaa
     3601 ggatcttcac ctagatcctt ttatattata aatgaagttt tatatcaatc taaagtatat
     3661 atgagtaaac ttggtctgac agttaccaat gcttgatcag tgaggcacct atctcagcga
     3721 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3781 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3841 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3901 caactttatc cgcctccatc cagtctatca attgttgccg ggaagctaga gtaagtagtt
     3961 cgccagtcaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4021 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4081 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4141 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4201 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4261 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4321 atagcagaac tttgaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4381 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4441 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4501 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4561 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4621 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcc
     4681 caatgatatc atttatatgt cgacattgat tattgactag ttattattag taatcaatta
     4741 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4801 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4861 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggactat ttacggtaaa
     4921 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     4981 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     5041 cttggcagta catctacgta ttagtcatcg ctattaccat gggtcgaggt gagccccacg
     5101 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     5161 ttttatttat tttgtgcagc gatgggggcg gggggggggg g
//
