LOCUS       sequence-308108-        5210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector based on the LiOn integration-coupled translational switch 
            (Kumamoto et al bioRxiv 2019) 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 5210)
  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 5210)
  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..5210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          658..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1739..1748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   1762..1796
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2127..2150)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2325..2344)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2594..2612
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     repeat_region   2730..2792
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     3148..3167
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3166..3221
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3267..3286
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     rep_origin      complement(3433..4021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3513..3532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4192..4983,4984..5052))
                     /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     4815..4834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5053..5157)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgccr gggggggggg gggggggggg gggggggggg
      541 ggggsgrggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      661 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      721 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      781 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      841 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
      901 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      961 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1081 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggrggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagcacgtga ggaccggtgc
     1741 caccatggcc tcctccctta accctagaaa gatagtctgc gtaaaattga cgcatgcatc
     1801 aattgtaact gcggtttaaa ctccccagca tgcctgctat tgtcttccca atcctccccc
     1861 ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag acaatgcgat
     1921 gcaatttcct cattttatta ggaaaggaca gtgagagtgg caccttccag ggtcaaggaa
     1981 ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcactaa agtcggagaa
     2041 ttcattcccg ggggaaaaag atctttaggc gccggtggag gggcggccct cggcgcgctc
     2101 gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tgatgtcggt
     2161 cttgtaggcg ccgggcagct gcacgggctt cttggccatg taggtggtct tgacctcggc
     2221 gtcgtagtgg ccgccgtcct tcagcttcag cctcatcttg atctcgccct tcagggcgcc
     2281 gtcctcgggg tacatccgct cggtggaggc ctcccagccc atggtcttct tctgcattac
     2341 ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc ttgtagatga actcgccgtc
     2401 ctgcagggag gagtcctggg tcacggtcac cacgccgcca tcctcgaagt tcatcacgcg
     2461 ctcccacttg aagccctcgg ggaaggacag cttcaagtag tcggggatgt cggcggggtg
     2521 cttcacgtag gccttggagc cgtactggaa ctgaggggac aggatgtccc aggcgaaggg
     2581 cagggggccg cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc
     2641 ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg gagccctcca tgcgcacctt
     2701 gaagcgcatg aactccttga tgacattaac cctagaaaga taatcatatt gtgacgtacg
     2761 ttaaagataa tcatgcgtaa aattgacgca tgtccggagc cgtagatatc ggaaagaaag
     2821 cggccgctgc aggtcgaggg atctccataa gagaagaggg acagctatga ctgggagtag
     2881 tcaggagagg aggaaaaatc tggctagtaa aacatgtaag gaaaatttta gggatgttaa
     2941 agaaaaaaat aacacaaaac aaaatataaa aaaaatctaa cctcaagtca aggcttttct
     3001 atggaataag gaatggacag cagggggctg tttcatatac tgatgacctc tttatagcca
     3061 acctttgttc atggcagcca gcatatgggc atatgttgcc aaactctaaa ccaaatactc
     3121 attctgatgt tttaaatgat ttgccctccc atatgtcctt ccgagtgaga gacacaaaaa
     3181 attccaacac actattgcaa tgaaaataaa tttcctttat tagccagaag tcagatgctc
     3241 aaggggcttc atgatgtccc cataattttt ggcagaggga aaaagatctc agtggtattt
     3301 gtgagccagg gcattggcca caccagccac caccttctga taggcagcct gcacctgagg
     3361 agtgaattag aagctttgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3421 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3481 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3541 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3601 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3661 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3721 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3781 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3841 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3901 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3961 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4021 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4081 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4141 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4201 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4261 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4321 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4381 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4441 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4501 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4561 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4621 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4681 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4741 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4801 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4861 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4921 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4981 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5041 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5101 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5161 atttccccga aaagtgccac ctgacgtccc aatgatatca tttaaatgtc
//
