LOCUS       sequence-308117-        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"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1732..1741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   1755..1789
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2120..2143)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2318..2337)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2587..2605
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     repeat_region   2723..2785
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     3139..3158
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3157..3212
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3258..3277
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     rep_origin      complement(3424..4012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3504..3523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4183..4974,4975..5043))
                     /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     4806..4825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5044..5148)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattattagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      421 tcccccccct ccccaccccc aattttgtat ttatttattt tttatttatt ttgtgcagcg
      481 atgggggcgg gggggggggg gncgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt attgacggct cgtttctttt ctgtggctgc gtgaaagcct
      841 tatagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttgt tgtgctgtct catcattttg gcaaagcacg tgaggaccgg tgccaccatg
     1741 gcctcctccc ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc atcaattgta
     1801 actgcggttt atactcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     1861 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     1921 cctcatttta ttaggaaagg acagtgagag tggcaccttc cagggtcaag gaaggcacgg
     1981 gggaggggca aacaacagat ggctggcaac tagaaggcac taaagtcgga gaattcattc
     2041 ccgggggaaa aagatcttta ggcgccggtg gagtggcggc cctcggcgcg ctcgtactgt
     2101 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca gcttgatgtc ggtcttgtag
     2161 gcgccgggca gctgcacggg cttcttggcc atgtaggtgg tcttgacctc ggcgtcgtag
     2221 tggccgccgt ccttcagctt cagcctcatc ttgatctcgc ccttcagggc gccgtcctcg
     2281 gggtacatcc gctcggtgga ggcctcccag cccatggtct tcttctgcat tacggggccg
     2341 tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg
     2401 gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac
     2461 ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg
     2521 taggccttgg agccgtactg gaactgaggg gacaggatgt cccaggcgaa gggcaggggg
     2581 ccgcccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc
     2641 tcgccctcga tctcgaactc gtggccgttc acggagccct ccatgcgcac cttgaagcgc
     2701 atgaactcct tgatgacatt aaccctagaa agataatcat attgtgacgt acgttaaaga
     2761 taatcatgcg taaaattgac gcatgtccgg agccgtagat atcggaaaga aagcggccgc
     2821 tgcaggtcga gggatcttca taagagaaga gggacagcta tgactgggag tagtcaggag
     2881 aggaggaaaa atctggctag taaaacatgt aaggaaaatt ttagggatgt tatagaaaaa
     2941 aataacacaa aacaaaatat aaaaaaaatc taacctcaag tcaaggcttt tctatggaat
     3001 aaggaatgga cagcaggggg ctgtttcata tactgatgac ctctttatag ccacctttgt
     3061 tcatggcagc cagcatatgg catatgttgc caaactctaa accaaatact cattctgatg
     3121 ttttatatga tttgccctcc catatgtcct tccgagtgag agacacaaaa aattccaaca
     3181 cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct caaggggctt
     3241 catgatgtcc ccataatttt tggcagaggg aaaaagatct cagtggtatt tgtgagccag
     3301 ggcattggcc acaccagcca ccaccttctg ataggcagcc tgcacctgag gagtgaatta
     3361 gaagctttga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3421 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3481 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3541 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3601 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3661 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3721 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3781 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3841 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3901 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3961 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt tagggatttt
     4081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatattata aatgaagttt
     4141 tatatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttgatcag
     4201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4321 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatca attgttgccg
     4441 ggaagctaga gtaagtagtt cgccagtcaa tagtttgcgc aacgttgttg ccattgctac
     4501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4801 acgggataat accgcgccac atagcagaac tttgaaagtg ctcatcattg gaaaacgttc
     4861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5101 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5161 aaaagtgcca cctgacgtcc caatgatatc atttatatgt c
//
