LOCUS       sequence-297941-        5201 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 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"
     promoter        complement(228..332)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        386..765
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        768..1043
                     /label=chicken beta-actin promoter
     intron          1044..2052
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1976..1998
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      2117..2126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2140..2174
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2505..2528)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2703..2722)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2972..2990
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     repeat_region   3108..3170
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     3524..3543
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3542..3597
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3643..3662
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     rep_origin      complement(3809..4397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3889..3908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4568..5201,1..158,159..227))
                     /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     join(5191..5201,1..9)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
       61 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
      121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
      181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
      241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
      301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
      361 gtcccaatga tatcatttaa atgtcgacat tgattattga ctagttatta atagtaatca
      421 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      481 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      541 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga ctatttacgg
      601 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      661 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      721 cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggtcg aggtgagccc
      781 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      841 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc gcgccaggcg
      901 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
      961 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     1021 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt gccccgctcc
     1081 gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     1141 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt
     1201 cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg
     1261 agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggcccgcgc
     1321 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcgtgtgcg
     1381 cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga ggggaacaaa
     1441 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc ggcggtcggg
     1501 ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc
     1561 ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt
     1621 gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg
     1681 cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg
     1741 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag ccgaaatctg
     1801 ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg ccggcaggaa
     1861 ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccatctcca
     1921 gcctcggggc tgccgcaggg ggacggctgc cttcgggggg gacggggcag ggcggggttc
     1981 ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc
     2041 tttttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca ttttggcaaa
     2101 gcacgtgagg accggtgcca ccatggcctc ctcccttaac cctagaaaga tagtctgcgt
     2161 aaaattgacg catgcatcaa ttgtaactgc ggtttaaact ccccagcatg cctgctattg
     2221 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2281 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gagagtggca
     2341 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2401 ggcactaaag tcggagaatt cattcccggg ggaaaaagat ctttaggcgc cggtggagtg
     2461 gcggccctcg gcgcgctcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
     2521 gtccagcttg atgtcggtct tgtaggcgcc gggcagctgc acgggcttct tggccatgta
     2581 ggtggtcttg acctcggcgt cgtagtggcc gccgtccttc agcttcagcc tcatcttgat
     2641 ctcgcccttc agggcgccgt cctcggggta catccgctcg gtggaggcct cccagcccat
     2701 ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt
     2761 gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc
     2821 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc
     2881 ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tactggaact gaggggacag
     2941 gatgtcccag gcgaagggca gggggccgcc cttggtcacc ttcagcttgg cggtctgggt
     3001 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga
     3061 gccctccatg cgcaccttga agcgcatgaa ctccttgatg acattaaccc tagaaagata
     3121 atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg tccggagccg
     3181 tagatatcgg aaagaaagcg gccgctgcag gtcgagggat cttcataaga gaagagggac
     3241 agctatgact gggagtagtc aggagaggag gaaaaatctg gctagtaaaa catgtaagga
     3301 aaattttagg gatgttaaag aaaaaaataa cacaaaacaa aatataaaaa aaatctaacc
     3361 tcaagtcaag gcttttctat ggaataagga atggacagca gggggctgtt tcatatactg
     3421 atgacctctt tatagccacc tttgttcatg gcagccagca tatggcatat gttgccaaac
     3481 tctaaaccaa atactcattc tgatgtttta aatgatttgc cctcccatat gtccttccga
     3541 gtgagagaca caaaaaattc caacacacta ttgcaatgaa aataaatttc ctttattagc
     3601 cagaagtcag atgctcaagg ggcttcatga tgtccccata atttttggca gagggaaaaa
     3661 gatctcagtg gtatttgtga gccagggcat tggccacacc agccaccacc ttctgatagg
     3721 cagcctgcac ctgaggagtg aattagaagc tttgagcaaa aggccagcaa aaggccagga
     3781 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3841 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3901 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3961 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4021 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4081 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4141 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4201 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4261 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4321 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4381 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4441 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4501 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4561 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4621 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4681 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4741 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4801 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4861 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4921 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4981 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5041 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5101 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5161 cgagttgctc ttgcccggcg tcaatacggg ataataccgc g
//
